<%BANNER%>

Non-coding RNA in Mammalian Development and Disease

Permanent Link: http://ufdc.ufl.edu/UFE0024187/00001

Material Information

Title: Non-coding RNA in Mammalian Development and Disease
Physical Description: 1 online resource (86 p.)
Language: english
Creator: O'Rourke, Jason
Publisher: University of Florida
Place of Publication: Gainesville, Fla.
Publication Date: 2009

Subjects

Subjects / Keywords: apoptosis, congenital, dicer, dmpk, dystrophy, hypotonia, microrna, muscleblind, myogenesis, myotonic, rna, six5
Genetics (IDP) -- Dissertations, Academic -- UF
Genre: Medical Sciences thesis, Ph.D.
bibliography   ( marcgt )
theses   ( marcgt )
government publication (state, provincial, terriorial, dependent)   ( marcgt )
born-digital   ( sobekcm )
Electronic Thesis or Dissertation

Notes

Abstract: The mammalian genome is a remarkable blueprint for a sophisticated and complex being consisting of trillions of cells that have the ability to rapidly respond to internal and external stimuli. During embryonic development, a single fertilized egg undergoes multiple cellular divisions while regulating a variety of biochemical processes inside the mother to morph into a physiologically complicated fetus that must be prepared for life outside the womb. Postnatally, thriving neonates continue to develop and adapt to a dynamic intracellular environment and external world. Although it is clear that an organism s DNA encodes its ability to develop and react to its surroundings, the precise mechanisms by which genes and gene products regulate morphogenesis and disease are only beginning to be uncovered. Recent mapping of functional sequence elements in the human genome has revealed that non-protein coding RNA (ncRNA) gene products are abundant and comprise a significant portion of the transcriptome. This study aims to examine fundamental aspects of ncRNA during mammalian development and to determine how mutations in ncRNA result in disease pathogenesis. We investigated the role of Dicer-dependent ncRNAs during skeletal muscle development by mutating Dicer specifically in the myogenic compartment during mouse embryogenesis. Our analysis demonstrates that Dicer-dependent ncRNAs are critical components required for normal muscle development during embryogenesis and postnatal life. Dicer mutant embryos display decreased skeletal muscle mass accompanied by abnormal myofiber morphology. Dicer mutants also show increased muscle apoptosis and reduced cell proliferation. Our findings demonstrate key roles for ncRNAs in mammalian development. We also took a disease orientated approach to determine how mutations in non-coding regions of mRNA cause congenital myotonic dystrophy (CDM), a disease characterized by delays in embryonic and postnatal development. CDM results from the expansion of a non-coding CTG repeat in the DMPK gene. Although molecular defects underlying CDM have not been uncovered, there is evidence that MBNL sequestration by toxic CUG repeat containing RNA and altered expression of DMPK-linked genes may be involved in pathogenesis. Here, we test the hypothesis that CDM results from MBNL1 loss combined with reduced expression of DMPK and/or the DMPK-linked gene SIX5. Knockout mice lacking Mbnl1 and/or Dmpk and Six5 were evaluated for embryonic developmental delays characteristic of CDM. Mbnl1;Dmpk and Mbnl1;Six5 double knockout mice do not recapitulate the delayed myofiber maturation and joint contractures observed in CDM. These observations argue against a role for DMPK and SIX5 haploinsufficiency in CDM pathogenesis and highlight the necessity for further investigation into identifying CDM molecular defects.
General Note: In the series University of Florida Digital Collections.
General Note: Includes vita.
Bibliography: Includes bibliographical references.
Source of Description: Description based on online resource; title from PDF title page.
Source of Description: This bibliographic record is available under the Creative Commons CC0 public domain dedication. The University of Florida Libraries, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.
Statement of Responsibility: by Jason O'Rourke.
Thesis: Thesis (Ph.D.)--University of Florida, 2009.
Local: Adviser: Swanson, Maurice S.
Electronic Access: RESTRICTED TO UF STUDENTS, STAFF, FACULTY, AND ON-CAMPUS USE UNTIL 2010-05-31

Record Information

Source Institution: UFRGP
Rights Management: Applicable rights reserved.
Classification: lcc - LD1780 2009
System ID: UFE0024187:00001

Permanent Link: http://ufdc.ufl.edu/UFE0024187/00001

Material Information

Title: Non-coding RNA in Mammalian Development and Disease
Physical Description: 1 online resource (86 p.)
Language: english
Creator: O'Rourke, Jason
Publisher: University of Florida
Place of Publication: Gainesville, Fla.
Publication Date: 2009

Subjects

Subjects / Keywords: apoptosis, congenital, dicer, dmpk, dystrophy, hypotonia, microrna, muscleblind, myogenesis, myotonic, rna, six5
Genetics (IDP) -- Dissertations, Academic -- UF
Genre: Medical Sciences thesis, Ph.D.
bibliography   ( marcgt )
theses   ( marcgt )
government publication (state, provincial, terriorial, dependent)   ( marcgt )
born-digital   ( sobekcm )
Electronic Thesis or Dissertation

Notes

Abstract: The mammalian genome is a remarkable blueprint for a sophisticated and complex being consisting of trillions of cells that have the ability to rapidly respond to internal and external stimuli. During embryonic development, a single fertilized egg undergoes multiple cellular divisions while regulating a variety of biochemical processes inside the mother to morph into a physiologically complicated fetus that must be prepared for life outside the womb. Postnatally, thriving neonates continue to develop and adapt to a dynamic intracellular environment and external world. Although it is clear that an organism s DNA encodes its ability to develop and react to its surroundings, the precise mechanisms by which genes and gene products regulate morphogenesis and disease are only beginning to be uncovered. Recent mapping of functional sequence elements in the human genome has revealed that non-protein coding RNA (ncRNA) gene products are abundant and comprise a significant portion of the transcriptome. This study aims to examine fundamental aspects of ncRNA during mammalian development and to determine how mutations in ncRNA result in disease pathogenesis. We investigated the role of Dicer-dependent ncRNAs during skeletal muscle development by mutating Dicer specifically in the myogenic compartment during mouse embryogenesis. Our analysis demonstrates that Dicer-dependent ncRNAs are critical components required for normal muscle development during embryogenesis and postnatal life. Dicer mutant embryos display decreased skeletal muscle mass accompanied by abnormal myofiber morphology. Dicer mutants also show increased muscle apoptosis and reduced cell proliferation. Our findings demonstrate key roles for ncRNAs in mammalian development. We also took a disease orientated approach to determine how mutations in non-coding regions of mRNA cause congenital myotonic dystrophy (CDM), a disease characterized by delays in embryonic and postnatal development. CDM results from the expansion of a non-coding CTG repeat in the DMPK gene. Although molecular defects underlying CDM have not been uncovered, there is evidence that MBNL sequestration by toxic CUG repeat containing RNA and altered expression of DMPK-linked genes may be involved in pathogenesis. Here, we test the hypothesis that CDM results from MBNL1 loss combined with reduced expression of DMPK and/or the DMPK-linked gene SIX5. Knockout mice lacking Mbnl1 and/or Dmpk and Six5 were evaluated for embryonic developmental delays characteristic of CDM. Mbnl1;Dmpk and Mbnl1;Six5 double knockout mice do not recapitulate the delayed myofiber maturation and joint contractures observed in CDM. These observations argue against a role for DMPK and SIX5 haploinsufficiency in CDM pathogenesis and highlight the necessity for further investigation into identifying CDM molecular defects.
General Note: In the series University of Florida Digital Collections.
General Note: Includes vita.
Bibliography: Includes bibliographical references.
Source of Description: Description based on online resource; title from PDF title page.
Source of Description: This bibliographic record is available under the Creative Commons CC0 public domain dedication. The University of Florida Libraries, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.
Statement of Responsibility: by Jason O'Rourke.
Thesis: Thesis (Ph.D.)--University of Florida, 2009.
Local: Adviser: Swanson, Maurice S.
Electronic Access: RESTRICTED TO UF STUDENTS, STAFF, FACULTY, AND ON-CAMPUS USE UNTIL 2010-05-31

Record Information

Source Institution: UFRGP
Rights Management: Applicable rights reserved.
Classification: lcc - LD1780 2009
System ID: UFE0024187:00001


This item has the following downloads:


Full Text





PAGE 1

! "! #$#%&$'(#)!*#+!(#!,+,,+-(+#! './.-$0,.#1!+#'!'(2.+2.! ! ! ! ! 34! 5+2$#!*$3.*1!$6*$7*8.! ! ! ! ! +!'(22.*1+1($#!0*.2.#1.'! 1$!19.!)*+'7+1.!2&9$$-! $:!19.!7#(/.*2(1;!$:!:-$* ('+!(#!0+*1(+-!:7-:(--,.#1! $:!19.!*.<7(*.,.#12!:$*!19.!'.)*..!$:! '$&1$*!$:!09(-$2$09;! 7#(/.*2(1;!$:!:-$*('+! =>>?!

PAGE 2

! =! ! ! ! ! ! @!=>>?!5ABCD!*CEFGH!$6*CIGJF!

PAGE 3

! K! ! ! ! ! ! 1C!L4!MCNODP!QAGFDHBR!PGA DSQAGFDHBR!EGCHTFGB!ADS!SCP! !

PAGE 4

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

PAGE 5

! _! 1+3-.!$:!&$#1.#12! QAPF +&8#$V-.'),.#12ZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZU -(21!$:!1+3-.2ZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ ZZZZZZZZZZ` -(21!$:!:()7*.2ZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ ZZZZZZZZZa!! +321*+&1ZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ ZZZZZZZZZZZZZZ?! &9+01.*!! "!'(&.*!'.0.#'.#1!DX*#+2!+*.!. 22.#1(+-!:$*!,72&-.!'./.-$0,.#1ZZZZ"" (DHGCSIXHOCDZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ ZZZZZZZZZZ"" #X*#+B!&CDBHOHIHF!,CBH!CW!HTF!9ILAD!)FDCLFZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ"" 'OBXCNFG4!$W!==!#IXMFCHOSF!bDHc!*FPIMAHCG4!DX*#+BZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ"= ,O*#+3OCPFDFBOB!ADS!,AHIGAHOCDZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ"= ,O*#+B!(DTOEOH!0GCHFOD! 24DHTFBOB!OD!,ALLAMBZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ"K ,OGDA!1AGPFHB!ADS!1AGPFH!0GFSOXHOCDZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ"U ,O*#+B!'OBQMA4!*FBHGOXHFS!2QAHOCHFLQCGAM!.]QGFBBOCDZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ"U ,O*#+!8DCXJCIH!,OXF!*FNFAM!2IEHMF !*CMFB!WCG!(DSONOSIAM!,O*#+BZZZZZZZZZZZZZZZZZZZZZZZZZZ"_ 'OXFG!,IHAHOCDB!)ONF!&MIFB! HC!,O*#+!)MCEAM!:IDXHOCDBZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ"_ *FBIMHBZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ ZZZZZZZZZZZZZ"d ,IBXMF!,O*#+!-FNFMB!+GF!*FSIXFS!:CMMC [ODP!'OXFG!(DAXHONAHOCD!OD!2JFMFHAM! ,IBXMFZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ Z"d 'OXFG!(DAXHONAHOCD!*FBIMHB!OD!0FGODAHAM!-F HTAMOH4!ADS!2JFMFHAM!,IBXMF!94QCQMABOAZZZZZZZZ"a 'FWFXHONF!,CGQTCPFDFBOB!ADS!,AODHFDA DXF!CW!'OXFG!,IHADH!,4CWOEFGBZZZZZZZZZZZZZZZZZZZZZZZ"? (DXGFABFS!+QCQHCBOB!OD! 'OXFG!,IHADH!,IBXMFZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ=> -CBB!CW!'OXFG!OD!,4CEMABHB!+MBC!-FASB!HC!.DTADXFS!&FMM!'FAHTZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ=" 'FXGFABFS!&FMM!0GCMOWFGAHOCD!:C MMC[ODP!-CBB!CW!,IBXMF!,O*#+BZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ=U ,4CPFDOX!*FPIMAHCG4!:AXHCGB!+GF!+EFGGADHM4 !.]QGFBBFS!AWHFG!'OXFG!(DAXHONAHOCDZZZZZZZZ=U 'OBXIBBOCDZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ ZZZZZZZZZZ=_ 'OXFG!OB!.BBFDHOAM!WCG!2JFMFHAM!,IBXMF!,CGQTCPFDFBOBZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ=_ 'OXFG!OB!*FYIOGFS!WCG!,IBXMF!&FMM!2IGNON AM!ADS!0GCMOWFGAHOCD!SIGODP!.LEG4CPFDFBOBZZZ=d! =!&$,3(#+1$*(+-!-$22!$:!,3#-"!+#'!',08%-(#8.'!).#.!.e0*.22($#! :+(-2!1$!,$'.-!&$#).#(1+-!,;$1$#(&!';21*$09;ZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZU> (DHGCSIXHOCDZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ ZZZZZZZZZZU> +SIMH%$DBFH!,4CHCDOX!'4BHGCQT4ZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZU> *#+!'CLODADXF!,CSFM!WCG!,4CHCDOX!'4BHGCQT4ZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZU" &CDPFDOHAM!,4CHCDOX!'4BHGCQT4!b&',cZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZU" 1TF! +,-' -CXIBf! +!.+ R! +!/0 !+DS! 1234 ZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZU= 1TF!'LQJ!.]QADSFS!&1)!*FQFAH!+MHFGB! 'L"!-CXIB!&TGCLAHOD!&CDWCGLAHOCDZZZZZZZZZZUU

PAGE 6

! d! 'OBGIQHOCD!CW!1TF!,CIBF!'L"!-CXIBZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZU_ *FBIMHBZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ ZZZZZZZZZZZZZU` #CGLAM!#FCDAHAM!0TFDCH4QF!WCG!,EDM"g 'LQJ!+DS!,EDM"g2O]_ !'CIEMF!8DCXJCIH! ,OXFZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ ZZU` &CLEODAHCGOAM!,EDM"R!'LQJ!ADS!2O]_!8 DCXJCIHB!:AOM!HC!,CSFM!&',%*FMFNADH! 'FMA4B!OD!,IBXMF!'FNFMCQLFDHZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZUa +EBFDXF!CW!-OLE!'FWCGLOHOFB!OD!-AHF!2HAPF!8DCXJCIH!.LEG4CBZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZU? 'OBXIBBOCDZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ ZZZZZZZZZZ_>! K!!!!!&$#&-7'(#)!*.,+*8 2!+#'!:717*.!'(*.&1($#2ZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZd"! U!,+1.*(+-2!+#'!,.19$'2ZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZdU ,OXF!)FDFGAHOCD!ADS!)FDCH4QODPZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZdU *#+!3MCHB!ADS!*1%0&*ZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZd_ 9OBHCMCP4R!e%PAM!2HAODODPR! (LLIDCTOBHCXTFLOBHG4!ADS!17#.-!+BBA4BZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZd` (LLIDCEMCHHODPZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ ZZZZZda ,CGQTCLFHGOX!ADS!2HAHOBHOXAM!+DAM4BOBZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZd? &FMM!&IMHIGFR!+DDF]OD%/%:(1&h0GCQOSOIL !(CSOSF!2HAODODP!ADS!&FMM!2CGHODPZZZZZZZZZZZZZZZZZZZZZZZZ`> 2JFMFHAM!QGFQAGAHOCDBZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ ZZZZZ`" -(21!$:!*.:.*.#&.2ZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ ZZ`= 3($)*+09(&+-!28.1&9ZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZad

PAGE 7

! `! -(21!$:!1+3-.2! 1AEMF !! QAPF !"%"!,O*#+B!OSFDHOWOFS!OD!A!NA GOFH4!CW!SOWWFGFDH!CGPADOBLBZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ=?

PAGE 8

! a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d!(DXGFABFS!AQCQHCBOB!OD! +56%&' LIHADH!BJFMFHAM!LIBXMFZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZKd "%`!'FXGFABFS!LIBXMF!LO*#+B! ADS!ODXGFABFS!AQCQHCBOB!CW! +56%& !LIHADH!LIBXMF!XFMMB! 57' 85#&" ZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ ZZZZZK` "%a!'FXGFABFS!XFMM!QGCMOWFGAHOCD!OD! +56%&' LIHADH!BJFMFHAM!LIBXMFZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZKa "%?! +56%&' LIHADHB!BTC[!AEFGGADH!F]QGFB BOCD!CW!L4CPFDOX!WAXHCGBZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZK? =%"!',"!ADS!',=!AGF!XAIBFS!E4!LOXGCBAHFMMO HF!F]QADBOCD!OD!DCD%XCSODP!GFPOCDB!CG! +!/0 !ADS! 9(:; R!GFBQFXHONFM4ZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ__ =%=!*#+!'CLODADXF!,CSFMZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ_d =%K!1TGFF!HOPTHM4%MODJFS!PFDFB!AH!HTF! +!!MCXIBZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ_` =%U!-AHF!BHAPF!FLEG4CB!MAXJODP!,EDM"! ADS!'LQJ!CG!,EDM"!ADS!2O]_!SC!DCH!BTC[! SFNFMCQLFDHAM!SFWFXHBZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ_a =%_!,4CWOEFG!SFNFMCQLFDH!OB!DCH!SFMA 4FS!OD!,EDMg'LQJ!CG!,EDMg2O]_!SCIEMF! JDCXJCIHBZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZZ ZZZZZ_? =%d!3CDF!SFWCGLOHOFB!AGF!DCH!CEBFGNFS!OD! ,EDMg'LQJ!CG!,EDMg2O]_!SCIEMF!JDCXJCIHBZZZZZZZZd>

PAGE 9

! ?! +EBHGAXH!CW!'OBBFGHAHOCD!0GFB FDHFS!HC!HTF!)GASIAHF!2XTCCM! CW!HTF!7DONFGBOH4!CW!:MCGOSA!OD!0AGHOAM!:IMWOMMLFDH!CW!HTF! *FYIOGFLFDHB!WCG!HTF!'FPGFF!CW!'CXHCG!CW!0TOMCBCQT4! #$#%&$'(#)!*#+!(#!,+,,+-(+#! './.-$0,.#1!+#'!'(2.+2.!! 34! 5ABCD!$6*CIGJF! ,A4!=>>?! &TAOGf!,AIGOXF!2[ADBCD! ,AiCGf!,FSOXAM!2XOFDXFB%%)FDFHOXB! !1TF!LALLAMOAD!PFDCLF!OB!A!GFLAGJAEMF!EMIFQGODH!WCG!A!BCQTOBHOXAHFS!ADS!XCLQMF]!EFODP! XCDBOBHODP!CW!HGOMMOCDB!CW!XFMMB!HTAH!TANF!HTF!AE OMOH4!HC!GAQOSM4!GFBQCDS!HC !ODHFGDAM!ADS!F]HFGDAM! BHOLIMOZ!!'IGODP!FLEG4CDOX!SFNFMCQLFDHR!A!BO DPMF!WFGHOMO\FS!FPP!IDSFGPCFB!LIMHOQMF!XFMMIMAG! SONOBOCDB![TOMF!GFPIMAHODP! A!NAGOFH4!CW!EOCXTFLOXAM!QGCXFBBFB!OD BOSF!HTF!LCHTFG!HC!LCGQT!ODHC!A! QT4BOCMCPOXAMM4!XCLQMOXAHFS!WFHIB!HTAH!LIBH!EF!QG FQAGFS!WCG!MOWF!CIHBOSF! HTF![CLEZ!!0CBHDAHAMM4R! HTGONODP!DFCDAHFB!XCDHODIF!HC!SFNFMCQ!ADS!AS AQH!HC!A!S4DALOX!ODHGAXF MMIMAG!FDNOGCDLFDH!ADS! F]HFGDAM![CGMSZ!!+MHTCIPT!OH!OB!XMFAG!HTAH!AD!CG PADOBL6B!'#+!FDXCSFB!OHB! AEOMOH4!HC!SFNFMCQ!ADS! GFAXH!HC!OHB!BIGGCIDSODPBR!HTF!QGFXOBF!LFXTADOB LB!E4![TOXT!PFDFB!ADS!PFDF!QGCSIXHB!GFPIMAHF! LCGQTCPFDFBOB!ADS!SOBFABF!AGF!CDM4!EFPODDODP!HC !EF!IDXCNFGFSZ!!*FXFDH!LAQQODP!CW!WIDXHOCDAM! BFYIFDXF!FMFLFDHB!OD!HTF!TILAD!PFDCLF!TAB!GF NFAMFS!HTAH!DCD%QGCH FOD!XCSODP!*#+!bDX*#+c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

PAGE 10

! ">! LIBXMF!SFNFMCQLFDH!SIGODP!FLEG4CPFDFBOB!ADS!QCBHDAHAM!MOWFZ!! +56%& !LIHADH!FLEG4CB!SOBQMA4! SFXGFABFS!BJFMFHAM!LIBXMF!LABB!AXXCLQ ADOFS!E4!AEDCGLAM!L4CWOEFG!LCGQTCMCP4Z!! +56%&' LIHADHB! AMBC!BTC[!ODXGFABFS!LIBXMF!A QCQHCBOB!ADS!GFSIXFS!XFMM!QGCMOWFG AHOCDZ!!$IG!WODSODPB!SFLCDBHGAHF! JF4!GCMFB!WCG!DX*#+B!OD!LALLAMOAD!SFNFMCQLFDHZ!!! VF!AMBC!HCCJ!A!SOBFABF!CGOFDHAHFS!AQQGCAXT !HC!SFHFGLODF!TC[!LIHAHOCDB!OD!DCD%XCSODP! GFPOCDB!CW!L*#+!XAIBF!XCDPFDOHAM!L4CHCDOX!S4 BHGCQT4!b&',c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hCG!HTF! +!/0 %MODJFS!PFDF! 1234 Z!!8DCXJCIH!LOXF!MAXJODP!,EDM"!ADShCG!'LQJ!ADS! 2O]_![FGF!FNAMIAHFS!WCG!FLEG4CDOX!SFNFMCQL FDHAM!SFMA4B!XTAGAXHFGOBHOX!CW!&',Z!! !<7=->+,?@ ADS! !<7=->15A4 !SCIEMF!JDCXJCIH!LOXF!SC!DCH!GFXAQOHIMAHF!HT F!SFMA4FS!L4CWOEFG!LAHIGAHOCD!ADS! iCODH!XCDHGAXHIGFB!CEBFGNFS!OD!&',Z!!1TFBF!CEB FGNAHOCDB!AGPIF!APAODBH!A!GCMF!WCG!',08!ADS! 2(e_!TAQMCODBIWWOXOFDX4!OD!&',!QAHTCPFDFBO B!ADS!TOPTMOPTH!HTF!DFXFBBOH4!WCG!WIGHTFG! ODNFBHOPAHOCD!ODHC!OSFDHOW4 ODP!&',!LCMFXIMAG!SFWFXHBZ !!!!!!!!!!!

PAGE 11

! ""! &9+01.*!"! '(&.*!'.0.#'.#1!DX*#+2!+*.!. 22.#1(+-!:$*!,72&-.!'./.-$0,.#1! Introduction NcRNAs Constitute Most of the Human Genome 1TF!XFDHGAM!SCPLA!CW!LCSFGD! EOCMCP4!BHAHFB!HTAH !QGCHFOD!B4DHTFBOB!GFBIMHB!WGCL!AD! OGGFNFGBOEMF!MODFAG!QGCXFBB!E4![TOXT!PFDFHOX!OD WCGLAHOCD!FDXCSFS!E4!'#+!OB!HGADBXGOEFS!ODHC! ODHFGLFSOAHF!LFBBFDPFG!*#+!bL*#+c!LCMFXIMFB!HTAH! AGF!HGADBMAHFS!ODHC!QG CHFOD!b"cZ!!#OGFDEFGPR! $XTCA!ADS!8TCGADA!SOBXCNFGFS!HTF!PFDFHOX!XCSF! OD!"?d"!ADS!IDHOM!GFXFDHM4!OH!TAB!EFFD!QGFBILFS! HTAH!QGCHFODB!AGF!HTF!LCMFXIMFB!GFBQCDBOEMF!WC G!BQFXOW4ODP!TILAD!WCGL! ADS!WIDXHOCDZ!!(DSFFSR!! QGCHFODB!AGF!F]YIOBOHF!LAXGCLCMFXIMFB!HTAH!TANF! EFFD!BTC[D!HC!TANF!XAH AM4HOX!ADS!GFPIMAHCG4! WIDXHOCDB![TOXT!FDAEMF!HTFL!HC!TANF!SONFGBF!A WWFXHB!CD!XFMMIMAG!QGCXFBBF BZ!!9C[FNFGR!CHTFG!DCD% QGCHFOD!LCMFXIMFB!TANF!GFXFDHM4!FLFGPFS!HTAH! GFPIMAHF!TILAD!QT4BOCMCP4Z!!DX*#+B!AGF!*#+! LCMFXIMFB!HTAH!AGF!HGADBXGOEFS!WG CL!A!'#+!HFLQMAHF!ADS!AGF!DCH!HGADBMAHFS!ODHC!QGCHFOD!b=cZ!! DX*#+B!AGF!PAODODP!PGFAHFG!BHAHIB!SIF!HC!GFXFD H!BHISOFB!SFLCDBHGAHODP! HTAH!AQQGC]OLAHFM4!`>% a>j!CW!HTF!TILAD!PFDCLF!OB!HGADBXGOEFSR!4FH!CDM4! "%=j!OB!HGADBMAHFS!ODHC !QGCHFOD!bKcZ!!1TOB! CEBFGNAHOCD!BIPPFBHB!HTAH!HTF!EIMJ!CW!HTF!TILAD!HG ADBXGOQHCLF!XCDBOBHB!CW!DX*#+Z!!1TOB!OB!OD! XCDHGABH!HC!MFBB!XCLQMF]!CGPADOBLB!BIXT!AB!EAXHFG OA!HTAH!HGADBMAHF!a>%?_j!CW!HTFOG!*#+!bUcZ!! :IGHTFGR!HTF!QGCHFCLFB!CW!BOLQMF!ADS!XCLQMF]!CG PADOBLB!AGF!BHGOJODPM4!BOLOMAGZ!!:CG!F]ALQMFR! TILADBR![TOXT!XCDBOBH!CW!HGOMMOCDB!CW!XFMMBR!TANF! QGCHFCLFB!HTAH!AGF!PGFAHF G!HTAD!`>j!OSFDHOXAM!HC! HTF![CGL! BC'%=%D)7E !HTAH!TAB!CDM4!k!"R>>>!BCLAHOX!XFMMBZ!! 1AJFD!HCPFHTFGR!HTFBF!CEBFGNAHOCDB! BIPPFBH!HTAH!DX*#+B!XCDHGOEIHF !HC!HTF!SFNFMCQLFDHAM!ADS!QT4BOCMCPOXAM!XCLQMF]OH4!ABBCXOAHFS! [OHT!TILADBZ! $DXF!HGADBXGOEFSR!DX*#+B!AGF!QGCXFBBFS!OD!BOL OMAG!WABTOCD!HC!L*#+BZ!!1TF4!AGF!XAQQFSR! QCM4ASFD4MAHFS!ADS!CWHFD!BQMOXFS!AD S!FSOHFS!b_cZ!!(D!XCDHGABH!HC !L*#+BR!DX*#+B!XAD!AMBC!EF!

PAGE 12

! "=! XMFANFS!ADS!QGCXFBBFS!ODHC!BLAMMF G!WIDXHOCDAM!IDOHB!bdcZ!!1TFGF! AGF!AMBC!SOWWFGFDH!XMABBFB!CW! DX*#+!b=cZ!!2CLF!TANF!TCIBF%JFFQODP!GCMFB!ADS! AGF!XCDBHOHIHONFM4!ADS!IE OYIOHCIBM4!F]QGFBBFSZ!! 1TFBF!ODWGABHGIXHIGAM!DX*#+B! ODXMISF!GOECBCLAM!*#+B!bG*#+BcR!HGADBWFG!*#+B!bH*#+Bc!ADS! BLAMM!DIXMFCMAG!*#+B!bBDC*#+Bc!HTAH!WIDXHOCD! SIGODP!HGADBMAHOCDR!ADS!BLAMM!DIXMFAG!*#+B! bBD*#+Bc!HTAH!GFPIMAHF!QGF%L*#+!BQMOXODPZ!!$H TFG!DX*#+B!BTC[!GFBHGOXHFS!BQAHOCHFLQCGAM! F]QGFBBOCD!ADS!TANF!FNCMNFS!LCGF!BQFXOAMO\FS! WIDXHOCDBZ!!:CG!F]ALQMFR!e(21!OB!A!DX*#+! ODNCMNFS!OD!e!XTGCLCBCLF!AXHONAHOCD!b`cR!ADS! LOXGC*#+B!bLO*#+Bc!AGF!A!SOWWFGFDH!XMABB!CW! DX*#+!HTAH!WIDXHOCD!QGOLAGOM4!HC!GFPIMAHF!PF DF!F]QGFBBOCD!QCBH%HGA DBXGOQHOCDAMM4!bacZ! Discovery of 22 Nucleotide (nt) Regulatory ncRNAs LOXGC*#+B!bLO*#+Bc![FGF!SOBXCNFGFS!AB!==! DH!DX*#+B!HTAH!XCDHGCM!SFNFMCQLFDHAM! HOLODP!OD! BC'%=%D)7E' b?cZ!!MOD%UR!HTF!WOGBH!LO*#+!XM CDFSR![AB!BTC[D!HC!GFPIMAHF!HTF! SFNFMCQLFDHAM!HGADBOHOCD!EFH[ FFD!-"!ADS!-=!BHAPFB!CW! BC%=%D)7E !MAGNAM!SFNFMCQLFDH!E4!QCBH% HGADBXGOQHOCDAMM4!GFPIMAH ODP!HTF!F]QGFBBOCD!CW! =57F-G' b">cZ!!2FNFD!4FAGB!MAHFG!HTF!BFXCDS!LO*#+R! MFH%`R![AB!OSFDHOWOFS!ADS!OH!HCC![AB!FBBFDHOAM!SIGOD P![CGL!MAGNAM!SFNFMCQLFDH!b""cZ!!1TF!WODSODP! HTAH!MFH%`![AB!FNCMIHOCDAGOM4!X CDBFGNFS!WGCL![CGLB!HC!LALLAMB!b"=c!MFS!HC!HTF!OSFDHOWOXAHOCD!CW! DILFGCIB!LO*#+B!OD![CGLBR!WMOFBR!\FEGAWOBTR!QMAD HB!ADS!LALLAMB!b"K%"?c!b1AEMF!"%"cZ!!:IGHTFGR! OH!OB!FBHOLAHFS!HTAH!HTFGF!AGF!HTCIBADSB!CW! LO*#+B!FDXCSFS!E4!HTF!TILAD!PFDCLF![TOXT!LA4! GFPIMAHF!=>%K>j!CW!JDC[D!PFDFB!b=>R="cZ! MiRNA Biogenesis and Maturation +MHTCIPT!WIDXHOCDAM!LO*#+B!AGF!="%=K!DH!MCDPR!HTF4 !CGOPODAHF!AB!HGADBX GOQHB!XCDBOBHODP!CW!! TIDSGFSB!HC!HTCIBADSB!CW!DIXMFCHOSFB!b:OPIGF! "c!b==cZ!!LO*#+B!AG F!MCXAHFS!HTGCIPTCIH!HTF! PFDCLF!ADS!TANF!EFFD!WCIDS!OD!F]CDBR!ODHGCDBR!ADS! ODHFGPFDOX!GFPOCDB!ADS!XAD!EF!WCIDS!AMCDF!CG! XMIBHFGFS!HCPFHTFG!b"aR=KcZ!!1T F4!AGF!HGADBXGOEFS!E4!*#+!QCM4LFGABF!((!ODHC!LCDCXOBHGCDOX!CG! QCM4XOBHGCDOX!QGOLAG4!bQGOc%LO*#+B !HTAH!WCGL!SCIEMF%BHGADSFS!TAOG QOD!BHGIXHIGFB!XCDHAODODP!A!_6!

PAGE 13

! "K! `%LFHT4M!PIADCBODF!XAQ!ADS!A!K6! QCM4%+!HAOM!b=UR=_cZ!!0GO%LO*#+B !AGF!QGCXFBBFS!OD!HTF!DIXMFIB! E4!HTF!SCIEMF%BHGADSFS!FDSCGOECDIXM FABF!'GCBTA!ODHC!k`>!DH!SCIEM F%BHGADSFS!QGFXIGBCG!bQGFc% LO*#+B![TOXT!AGF!F]QCGHFS!HC!HTF!X4HCQMABL! E4!.]QCGHOD!_!b=d%=acZ!!0GF%LO*#+B!HTFD!IDSFGPC! A!WODAM!XMFANAPF!GFAXHOCD!E4!ADCHTFG!*#ABF!(((!FD SCDIXMFABFR!'OXFGR!HC!PFDFGAHF!SCIEMF%BHGADSFS! ="%=K!DH!*#+!LCMFXIMFB!b=`cZ!!2IEBFYIFDHM4R!HTFB F!BTCGH!SB*#+B!AGF!IDGANFMFS!E4!A!TFMOXABF!HC! PFDFGAHF!LAHIGF!==!DH!WIDXHOCDAM!LO *#+B!OD!HTF!XFMM!X4HCQMABL!b=?cZ! MiRNAs Inhibit Protein Synthesis in Mammals (D!HTF!X4HCQMABLR!HTF!==!DH!LO*#+!ODXCGQCGAH FB!ODHC!HTF!GOECDIXMF CQGCHFOD!XCLQMF]!JDC[D! AB!HTF!*#+%ODSIXFS!BOMFDXODP!XCLQMF]!b*(2&c! bK>cZ!!*(2&!OSFDHOWOFB!HAGPFH!LFBBFDPFG!*#+B! bL*#+Bc!EABFS!CD!XCLQMFLFDHAGOH4!EFH[FFD!HTF! LO*#+!ADS!OHB!HAGPFHZ!!(H![AB!CGOPODAMM4! QGCQCBFS!HTAH!HTF!SFPGFF!CW!XCLQMFLFDHAG OH4!SFHFGLODFS!HTF!L FXTADOBL!E4![TOXT!HTF! LO*#+h*(2&!GFQGFBBFS!HGADBMAHOCD! CW!HTF!HAGPFH!L*#+!bK"%KUcZ!!+ XXCGSODP!HC!HTOB!LCSFMR!OW!HTF! LO*#+!ADS!OHB!HAGPFH!AGF!QFGWFXHM4 !XCLQMFLFDHAG4!HTFD!HTF!L*#+!OB!SFPGASFSZ!!(W!HTFGF!OB!CDM4! QAGHOAM!XCLQMFLFDHAGOH4!HTF!LFBBAPF!OB!BHAEMF!EIH! OB!DCH!HGADBMAHFSZ!!.NOSFDXF!APAODBH!HTOB!LCSFM! XALF!WGCL!F]QFGOLFDHB![TOXT!BTC[FS!HTAH!QAGHOAM!XCLQMFLFDHAGOH4!EFH[FFD! =%#FH !LO*#+!ADS! OH6B!HAGPFH! =57FG!GFBIMHFS!OD!SF PGASAHOCD!CW! =57FG!LFBBAPF!OD! BC'%=%D)7E' bK_cZ!!1TOB!GAOBFS!HTF! QCBBOEOMOH4!HTAH!ADCHTFG!LFXTADOBLR!CG!WAXHCGR!SF HFGLODFB!OW!A!LO*#+h*(2& !SFPGASFB!CG!ODTOEOHB! HTF!HGADBMAHOCD!CW!HTF!L*#+Z!!L*#+!SFPGASAHO CD!CXXIGB!E4!HTF!BMOXFG!AXHONOH4!CW!+GPCDAIHF=R! CDF!CW!HTF!*(2&%ABBCXOAHFS!WAXHCG B!bKdcZ!!+GPCDAIHF=!XMFANFB!L*#+B! AH!A!BOHF!DFAG!HTF!LOSSMF!CW! HTF!LO*#+fL*#+!T4EGOS!bK`cZ!!1TF!QGFXOBF! LFXTADOBL!CW!HGADBMAHOCDAM!GFQGFBBOCD!E4! LO*#+h*(2&!OB!IDJDC[DZ!!9C[FNFGR!XAQ%SFQFDSFDH! HGADBMAHOCD!ODOHOAHOCD!CW !HTF!HAGPFHFS!L*#+! OB!ODTOEOHFS!ADS!HTFGF!OB!FNOSFDXF!HTAH!LO* #+h*(2&!SCXJODP!CD!L*#+!K671*B!QGFNFDHB!L*#+! XOGXIMAGO\HOCD!E4!ODHFGWFGODP![OHT!ODHFGAXHOCDB!EF H[FFD!WAXHCGB!HTAH!EODS!HTF!L*#+!_6!ADS!K6!FDSB! bKaRK?cZ!!

PAGE 14

! "U! MiRNA Targets and Target Prediction (D!CGSFG!HC!IDSFGBHADS!HTF!EOCM CPOXAM!GCMFB!CW!LO*#+BR!HTF!L*#+B!HTF4!HAGPFH!LIBH!EF! OSFDHOWOFSZ!!3OCODWCGLAHOX!AQQGC AXTFB!TANF!GFBIMHFS!OD!HTF!QGFSOXHOCD!CW!HTCIBADSB!CW!LO*#+! HAGPFHBZ!!1TFBF!AMPCGOHTLB!QGFSOXH!HAGPFHB!EABFS!CD !NAGOCIB!WFAHIGFB!CW!HT F!LO*#+!CG!HTF!HAGPFHR! BIXT!AB!HTF!_6%FDS!DIXMFCHOSF!B FFS!CW!HTF!LO*#+R!FNCMIHOCDAG4 !XCDBFGNAHOCD!EFH[FFD!BQFXOFBR! ADS!HTFGLCS4DALOX!WGFF!FDFGP4!XAMXIMAHOCDB!bU>%U`cZ !!+MHTCIPT!HTCIBADSB!CW!HAGPFHB!TANF!EFFD! QGFSOXHFS! 57'E5=56" R! 57'858"' HAGPFH!OSFDHOWOXAHOCD!GFLAODB!A!XTAMMFDPFZ!!3OCMCPOXAMM4!GFMFNADH!HAGPFHB! AGF!SOWWOXIMH!HC!OSFDHOW4!EFXAIBF!HTF!QGFXOBF!EOCX TFLOXAM!LFXTADOBL!CW!LO *#+h*(2&!HAGPFHODP!CW! L*#+B!OB!QCCGM4!IDSFGBHCCSZ!!:IGHTF GR!OH!OB!DCH!XMFAG!OW!HTF! 57'85#&" !K6!71*!GFQCGHFG!ABBA4B! XIGGFDHM4!IBFS!HC!NAMOSAHF!LO*#+!HAGPFHB!LOLOX! 57'858"' GFPIMAHOCDZ!!)AODODP!A!EFHHFG! IDSFGBHADSODP!CW!LO*#+!HAGPFHODP! EOCXTFLOBHG4!XCLEODFS![OHT! HTF!SFNFMCQLFDH!CW!AXXIGAHF! TOPT!HTGCIPTQIH!HAGPFH!NAMOS AHOCD!HFXTDOYIFB!OB!GFYIOGF S!HC!SFHFGLODF!OW!HTF! 57'E5=56" !QGFSOXHFS! HAGPFHB!AGF!EOCMCPOXAMM4!GFMFNADHZ! MiRNAs Display Restricted Spatiotemporal Expression LO*#+B!BTC[!HOBBIF%BQFXOWOX!F] QGFBBOCD!QAHHFGDB!OD!WOBTR!LO XF!ADS!TILADBZ!!VFODTCMSB!FH! AMZ!IBFS!MCXJFS%DIXMFOX!AXOS%LCSOWOFS!CMOPCDIX MFCHOSF!QGCEFB!HC!SFHF GLODF!HTF!BQAHOAM!ADS! HFLQCGAM!F]QGFBBOCD!QAHHFGD!CW!""_!XCDBFGNFS!NFGH FEGAHF!LO*#+B!OD!\FEGAWOBT!FLEG4CB!bUacZ!! 1TF4!GFQCGHFS!HTAH!HTF!LAiCGOH4!CW!HTFBF!LO*# +B![FGF!CEBFGNFS!HC!EF !F]QGFBBFS!OD!A!TOPTM4! HOBBIF%BQFXOWOX!LADDFG!SIGODP!SFNF MCQLFDHZ!!:CG!F]ALQMFR!LO*%=>d! [AB!BQFXOWOXAMM4!F]QGFBBFS!OD! \FEGAWOBT!LIBXMFBZ!!(D!ASSOHOCDR!2FLQFGF!FH!AMZ!IB FS!#CGHTFGD!EMCHB!HC!XT AGAXHFGO\F!HTF!F]QGFBBOCD! CW!""?!QGFNOCIBM4!GFQCGHFS!LO* #+B!OD!ASIMH!CGPADB!WGCL!LOXF! ADS!TILADB!bU?cZ!!1TFOG!GFBIMHB! BTC[FS!HTAH!K>!CW!HTFBF!LO*#+B![F GF!BQFXOWOXAMM4!F]QGFBBFS!CG!PG FAHM4!FDGOXTFS!OD!A!QAGHOXIMAG! CGPADZ!!!LO*%"R!LO*%"KK!ADS!LO*%=>d![FGF!F]AL QMFB!CW!LO*#+B!F]QGFBBFS!BQFXOWOXAMM4!OD!

PAGE 15

! "_! BJFMFHAM!LIBXMF!ADS!TFAGH!CW!ECH T!LOXF!ADS!TILADBZ!!1TFBF!BHISOF B!BIPPFBH!HTAH! SOWWFGFDH!LO*#+! PGCIQB!LA4!WIDXHOCD!HC!GFPIMAHF!XCDBFGNFS! SFNFMCQLFDHAM!CG!QT4BOCMCPOXAM!QAHT[A4BZ! MiRNA Knockout Mice Reveal Subt le Roles for Individual MiRNAs 'OWWFGFDH!PGCIQB!TANF!IBFS!A!GFNFGBF!PFDFHOXB !AQQGCAXT!HC!IDSFGBHADS!LO*#+!WIDXHOCD!OD! LALLAMBZ!!1C!SAHFR!WONF!LO*#+!JDCXJCIH!LCIBF!MOD FB!TANF!EFFD!PFDFGAHFS!b_>cb_"cb_=cb_KcZ!! (DHFGFBHODPM4R!AMM!LO*#+!JDCXJCIH!LOXF!AGF!NOAEMF !ADS!BTC[!LOMS!QTFDCH4QFBZ!!:CG!F]ALQMFR! HAGPFHFS!SFMFHOCD!CW!LO*%=>aR!A!TFAGH%BQFXOWOX!LO *#+R!SCFB!DCH!AQQFAG!HC!TANF!AD4!FWWFXH!CD! DCGLAM!XAGSOAX!WIDXHOCD!ADS!OH!OB!DCH!IDHOM!LO*%=>a!LIHA DH!TFAGHB!AGF!BHGFBBFS!HTAH!A!GCMF!WCG!LO*% =>a!OD!XAGSOAX!GFLCSFMODP!OB!CEBFGNFS!b_=cZ!!(D !ASSOHOCDR!LO*%"%=R!ADCH TFG!LO*#+!BQFXOWOXAMM4! FDGOXTFS!OD!LIBXMFR!JDCXJCIH!LOXF![FGF!DCH!CEBFG NFS!HC!TANF!AD4!BJFMFHAM!LIBXMF!QTFDCH4QF!ADS! TAS!CDM4!LODCG!SFWFXHB!OD!TFAGH!LCGQTCPFDFBOB! ADS!FMFXHGOXAM!XCDSIXHO CD!b_>cZ!!1TF!LOMS! QTFDCH4QF!ABBCXOAHFS![OHT!LO*%"%=!LIHADHB!XCIMS!EF! AHHGOEIHFS!OD!QAGH!HC!A!XCLQFDBAHCG4!AWWFXH! E4!LO*%"%"R!ADCHTFG!LO*%"!WALOM4!LFLEFG!HTAH!OB! F]QGFBBFS!AH!A!SOWWFGFDH!PFDFHOX!MCXIBZ!!1TF! BIEHMF!QTFDCH4QFB!CEBFGNFS!OD!LO*#+!JDCXJCIH! BHISOFB!BIPPFBH!HTAH!LO*#+B!LA4!BTAGF! WIDXHOCDAM!GFSIDSADX4!CG!TANF!BIEH MF!FWWFXHB!CD!XFMMIMAG! QGCXFBBFBZ!!(DSFFSR!H[C!GFXFDH!BHISOFB! BTC[FS!HTAH!ODSONOSIAM!LO*#+B!WIDXHOCD!AB!GTFC BHAHB!HC!WODF%HIDF!QGCHFOD!CIHQIH!b_UR__cZ!!!! Dicer Mutations Give Clues to MiRNA Global Functions (D!CGSFG!HC!SFHFGLODF!HTF!PMCEAM!WIDXHOCDB!CW! LO*#+BR!BFNFGAM!MAECGAHCGOFB!LIHAHFS!'OXFGR! HTF!FDSCDIXMFABF!GFBQCDBOEMF!WCG!QG CXFBBODP!QGF%LO*#+B!ODHC!HTFOG!WIDXHOCDAM!WCGLBZ!!-CBB!CW! 'OXFG!OD!HOBBIFB!ADS!XFMMB!GFBIMHB!OD!HTF!AXXILIMA HOCD!CW!HTF!QGF%LO*#+!ADS!MCBB!CW!HTF!WIDXHOCDAM! ==!DH!LO*#+Z!!,OXF!HTAH!MAXJ!'O XFG!SOF!AH!FLEG4CDOX!SA4!`Z_!b _dcZ!!:IGHTFGR!'OXFG!SFWOXOFDH! FLEG4CDOX!BHFL!XFMMB!TANF!QGCMOWFGAHOCD!SFWFXHB!A DS!ODXGFABFS!F]QGFBBOCD!WGCL!XFDHGCLFGFB!b_`cZ!! 'OXFG!TAB!AMBC!EFFD!XCDSOHOCDAMM4!ODAXHONAHFS!OD! A!NAGOFH4!CW!LCIBF!HOBBIFB!ODXMISODP!MOLEB!b_acR! BJOD!b_?cR!GFHODA!bd>cR!BQFGL!b d"cR!MIDP!bd=cR!WFLAMF!GFQGCSIXHON F!HGAXH!bdKcR!TFAGH!b_>cR!ADS!

PAGE 16

! "d! NAGOCIB!GFPOCDB!CW!HTF!XFDHGAM!DF GNCIB!B4BHFL!bdU%ddcZ!!(D!AMM!XAB FBR!'OXFG!SFWOXOFDH!LOXF!TANF! GFSIXFS!LO*#+!MFNFMB!ADS!LCBH!'OXFG!LIHADH B!TANF!A!NAGOFH4!CW!SFNFMCQLFDHAM!ADShCG! QT4BOCMCPOXAM!AEDCGLAMOHOFB!OD!HTF!HOBBIF![TFG F!'OXFG![AB!ODAXHONAHFSZ!!1TFGF![AB!AMBC!A! XCDBOSFGAEMF!ALCIDH!CW!XFMM!SFAH T!OD!BFNFGAM!SOWWFGFDH!'OXFG!JD CXJCIH!MODFBZ!!1TIBR!HTF!'OXFG! LIHADH!BHISOFB!SFLCDBHGAHF!HTAH!'OXFG!ADS!LO*# +B!AGF!FBBFDHOAM!WCG!QGCQFG!HOBBIF!LCGQTCPFDFBOB! ADS!BIPPFBH!HTAH!LO*#+B!LA4!EF! DFPAHONF!GFPIMAHCGB!CW!XFMM!SFAHTZ! Results Muscle MiRNA Levels are Reduced Following Dicer Inactivation in Skeletal Muscle 1C!E4QABB!HTF!FAGM4!FLEG4CDOX!MFHTAMOH4!ABBCXOAHFS ![OHT!A!'OXFG!DIMM!AM MFMF!ADS!ODNFBHOPAHF! OHB!GCMF!OD!BJFMFHAM!LIBXMFR![F!ODAXHONAHFS!'O XFG!BQFXOWOXAMM4!OD!BJFMFHAM!LIBXMF!IBODP!LOXF! XCDHAODODP!A!'OXFG!XCDSOHOCDAM!AMMFMF!ADS!A! ,4C'XGF!HGADBPFDF!b_aRd`cZ!1TF!'OXFG!XCDSOHOCDAM! AMMFMF!IBFS!OD!HTOB!BHIS4!TAB!HTF!F]CD!FDXCSO DP!CDF!CW!HTF!*#ABF!(((!SCLAODB!WMADJFS!E4!MC]0! BOHFB!b:OPZ!"%=+cZ!!&GF%LFSOAHFS!GFXCLEODAHOCD!CW !HTOB!F]CD!TAB!EFFD!BTC[D!HC!EF!FWWFXHONF!OD! GFSIXODP!'OXFG6B!AEOMOH4!HC!QGCXFBB!LO*#+B!ODHC !HTFOG!WIDXHOCDAM!WCGLB!b_acZ!!1TF!,4C'XGF! HGADBPFDF!XCDHAODB!&GF!GFXCLEODABF!SC[DBHGFAL!CW !,4C'!XCGF!FDTADXFG!ADS!QGCLCHFG!FMFLFDHB! b:OPZ!"%=+cZ!!,4C'XGF!AXHONOH4! OB!XCDBOBHFDH![OHT!FDSCPFDCIB!, 4C'!L*#+!F]QGFBBOCD!bd`cZ!!(H! OB!AXHONF!OD!HTF!LADSOEIMAG!AGXT!AH!.?Z`_R!BCL OHOX!L4CHCLFB!AH!.">Z_R!SFNFMCQODP!MOLEB!EISB!AH! .""Z_!ADS!AMM!HGIDJ!LIBXIMAHIGF!E4!."=Z_Z!!,OXF!X CDHAODODP!H[C![OMS!H4QF!AMMFMFBR!'OXFG! XCDSOHOCDAM!ADS![OMS!H4QF!AMMFMFB!bXhlcR!XCDSOHO CDAM!ADS!SFMFHOCD!AMMFMFB! bXh%c!CG!XCDSOHOCDAM!ADS! [OMS!H4QF!AMMFMFB!HCPFHTFG![OHT!,4C'XGF!bXhl g!,4C'XGFc![FGF!QTFDCH4QO XAMM4!ODSOBHODPIOBTAEMF! WGCL!CDF!ADCHTFG!ADS!AGF!GFWFGGFS!HC!AB!XCDHGC MBZ!!,OXF!XCDHAODODP!'OXFG!XCDSOHOCDAM!ADS!SFMFHOCD! AMMFMFB!XCLEODFS![OHT!,4C'XGF!bXh%g!,4C' XGFc!AGF!SFBOPDAHFS!AB!LIHADHBZ!!! VF!ABBFBBFS!'OXFG!ODAXHONAHOCD!OD!."`Z_!HCDPIF !LIBXMF!E4!*1%0&*![OHT!QGOLFGB!WMADJODP! WMC]FS!F]CD!=U!b:OPZ!"%=3cZ!!1TF!DFCL4XOD!GFBOBHAD XF!XABBFHHF!OB!OD!ODHGCD!=U!ADS![CIMS!DCH!TANF!

PAGE 17

! "`! EFFD!ALQMOWOFSZ!!1TF!LAiCGOH4!CW!'OXF G!HGADBXGOQHB!XCDHAODFS!F]CD!=U!OD!'OXFG Xhl !HCDPIF![TOMF! HTCBF!MAXJODP!F]CD!=U![FGF!QGFBFDH!AH!LIXT!MC[F G!MFNFMB!bMADF!"cZ!!1TF!AEBFDXF!CW!F]CD!=U! XCDHAODODP!L*#+!LA4!EF!F]QMAODF S!E4!MC[!MFNFM!AMHFGDA HONF!BQMOXODPZ!!1TOB! BIPPFBHB!HTAH!'OXFG! OBCWCGLB!MAXJODP!HTF!F]CD!=U!FDXCSFS!*#+BF!(((!SCLAOD!LOPTH!EF!GFYIOGFS!WCG!QGCQFGHOFB!CHTFG! HTAD!LO*#+!QGCXFBBODPZ!!+B!F]QFXHFSR!'OXFG Xh% !HCDPIF!LIBXMF!BTC[FS!GFMAHONFM4!FYIAM!ALCIDHB! CW!'OXFG!L*#+!XCDHAODODP!ADS!LO BBODP!F]CD!=U!bMADF!=cZ!!'OXFG Xh% g!,4C'XGF!LIHADH!BJFMFHAM! LIBXMF!SOBQMA4FS!TOPTFG!'OXFG!HG ADBXGOQH!MFNFMB!HTAH !MAXJFS!WMC]FS!F]CD!=U!bMADF!Kc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dR!IBODP![TCMF!MOLE!*#+!WGCL!."KZ_!ADS!. "UZ_!FLEG4CB!b:OPZ!"%=&cZ!!!LO*%"!MFNFMB! [FGF!GFSIXFS!E4!_=j!AH!."KZ_!ADS!`dj!AH!."UZ_! OD!LIHADHB!XCLQAGFS!HC!XCDHGCMBR!ADS!LO*%"KK! ADS!LO*%=>d!*#+B!FAXT!SFXGFABFS!E4!`Uj!AH!."KZ_! ADS!?>j!AH!."UZ_Z!!2OLOMAG!GFSIXHOCDB![FGF! CEBFGNFS!OD!LIHADHB!AH!MAHFG!SFNFMCQLFDHAM!BHAPFB !bSAHA!DCH!BTC[DcZ!!(D HFGFBHODPM4R!HTF!LAHIGF! LO*#+!MFNFM!GFSIXHOCD![AB!LCBH!SGALAHOX!AH! ."UZ_R!H[C!SA4B!AWHFG!,4C'XGF!OB!AXHONAHFS! HTGCIPTCIH!.!"=Z_!MOLE!LIBXMFZ!!1TOB!SFMA4!TAB!EFFD !CEBFGNFS!OD!CHTFG!HOBBIFB!ADS!XFMMB![TFGF! 'OXFG![AB!ODAXHONAHFS!ADS!LCBH! MOJFM4!GFWMFXHB!HTF!MCDP!TAMW%MOWF! CW!'OXFG!ADShCG!LO*#+Bb_?cZ!! 1TF!GFBOSIAM!ALCIDH!CW!WIDXHOCDA M!LO*#+!QGFBFDH!OD!LIHADH!MOLEB!LA4!TANF!GFBIMHFS!WGCL! ODXCLQMFHF!&GF%LFSOAHFS!GFXCLEODAHOCD!CG!WG CL!QCBBOEMF!F]QGFBBOCD!CW!HTFBF!LO*#+B!OD!DCD% LIBXMF!XFMM!H4QFB!OD!MOLEBR!ODXMISODP!ECDF!ADS!XAG HOMAPFZ!!1TFBF!GFBIMHB!SFLCDBHGAHFS!HTAH!LOXF!

PAGE 18

! "a! XCDHAODODP!'OXFG!XCDSOHOCDA M!AMMFMFB![OHT!A!,4C'XG F!HGADBPFDF!BTC[FS!AD!ODXGFABF!OD!'OXFG!F]CD! =U!SFMFHFS!HGADBXGOQHB![TOXT!GFBIMHFS!OD!S FXGFABFS!BJFMFHAM!LIBXMF!LO*#+!MFNFMBZ!! Dicer Inactivation Results in Perinatal Leth ality And Skeletal Muscle Hypoplasia +MM!'OXFG!BJFMFHAM!LIBXMF!LIHADHB!SOFS![ OHTOD!LODIHFB!WCMMC[ODP!EOGHTZ!!1TFGFWCGFR! QTFDCH4QOX!ADAM4BOB![AB!QFGWCGLF S!CD!DF[ECGDB!ADS!FLEG4CBZ!!VF!CEBFGNFS!HTAH!HTF!PGCBB! ADAHCL4!CW!MAHF!BHAPF!LIHADH!FLEG4CB!SOWWFGFS!WG CL!HTAH!CW!XCDHGCM!MOHHFGLAHFBZ!!2QFXOWOXAMM4R! LIHADH!FLEG4CDOX!MOLEB!AQQFAGFS! HC!EF!MFBB!SFNFMCQFS!b:OPZ!"%K+cZ!!1C!NOBIAMO\F!LIBXMF! LCGQTCMCP4R![F!e%PAM!BHAODFS!FLEG4CB!ADS!DF[ECG DB!XCDHAODODP!*$2+=d!GFQCGHFGB!bdac!HC! BQFXOWOXAMM4!BHAOD!HTCBF!XFMMB!OD![TOXT!&GF![AB!AXH ONF!ADS!MAXm![AB!F]QGFBBFS!b:OPZ!"%K3cZ!!VF! WCIDS!HTAH!LIHADH!FLEG4CB!BTC[FS!A!SFXGFABF! OD!LIBXMF!LABB!OD!ECHT!MOLEB!ADS!HCDPIFZ!! 9OBHCMCPOXAM!F]ALODAHOCD!CW!9n.!ADS!ADHO%SFB LODR!A!QAD!L4CWOEFG!LA GJFGR!BHAODFS!SOBHAM! WCGFMOLE!XGCBB!BFXHOCDB!XCDWOGLFS!HTAH!HTFGF![AB! MFBB!BJFMFHAM!LIBXMF!WCG!AMM!LIBXMFB!CW!LIHADH! FLEG4CDOX!MOLEB!b:OPZ!"%K&R!:OPZ!"%U+cZ!!1TOB! SFXGFABF![AB!WOGBH!CEBFGNFS!OD!."UZ_!LIHADH! FLEG4CB!ADS!EFXALF!LCGF!QGCDCIDXFS!SIGODP!BIEBF YIFDH!BHAPFB!CW!FLEG4CPFDFBOBZ!!34!."aZ_! HTF!ALCIDH!CW!LIBXMF!OD!LIHADH!MOLEB![AB!SGABHOXA MM4!GFSIXFS!XCLQAGFS!HC! XCDHGCM!MOHHFGLAHFBZ!! :IGHTFGLCGFR!A!QAHHFGDODP!SFWFXH![AB!DCH!CEBFGNF S!OD!LIHADH!FLEG4CB!AB!FAXT!CW!HTF!ODSONOSIAM! LIBXMFB![AB!QGFBFDH!AMEFOH! OD!BLAMMFG!QGCQCGHOCDBZ!!! VF!DF]H!BCIPTH!HC!SFHFGLODF!OW!HTF!GFSIXHOCD!OD! BJFMFHAM!LIBXMF![AB!SIF!HC!BJFMFHAM!LIBXMF! T4QCQMABOA!bWF[FG!WOEFGBc!ADShCG!T4QCHGCQT4!bBLAMMFG! WOEFGBcZ!!1C!SOBHODPI OBT!EFH[FFD!HTFBF!H[C! QCBBOEOMOHOFBR![F!XCIDHFS!HTF!DIL EFG!CW!L4CWOEFGB!QFG!PONFD!AGFA! ADS!LFABIGFS!L4CWOEFG!XGCBB% BFXHOCDAM!AGFA!WCG!FAXT!SOBHAM!MOLE!LIBXMF!CW!. "dZ_!FLEG4CB!b:OPZ!"%U3R!&cZ!!1TFBF!LFABIGFLFDHB! BTC[FS!HTAH!AMM!LIHADH!MOLE!LIBX MFB!XCDHAODFS!WF[FG!WOEFGB!ADS! HTAH!HTF!LIHADH!WOEFGB![FGF! MAGPFG!HTAD!HTF!XCDHGCMBZ!!+D!CNFGAMM!ODXGFABF! OD!ANFGAPF!L4CWOEFG!BO\F![AB!CEBFGNFS!EFXAIBF! LIHADH!LIBXMFB!XCDHAODFS!WF[FG!BLAMMFG!WOEFGB!XC LQAGFS!HC!XCDHGCMBZ!!VF!AMBC!F]ALODFS!WOEFG!

PAGE 19

! "?! H4QF!SOBHGOEIHOCD!OD!XCDHGCM!A DS!LIHADH!MOLEB!E4!OLLIDCTOBHCXTFLOBHG4![OHT!ADHOECSOFB!BQFXOWOX! WCG!L4CBOD!OBCWCGLB!QGFBFDH!OD!WABH!CG!BMC[!WO EFGBZ!!VF!CEBFGNFS!HTF!QGFBFDXF!CW!ECHT!WABH!ADS! BMC[!WOEFGB!ADS!BOLOMAG!BHAODODP!QAHHFGDB!OD!SOBH AM!WCGFMOLEB!CW!XCDHGCM!ADS!LIHADH!FLEG4CB!AH! ."KZ_%."dZ_!bSAHA!DCH!BTC[DcZ!!1TFBF!SAHA!BIPPFB HFS!HTAH!HTF!GFSIXHOCD!OD!LIBXMF!LABB!CEBFGNFS! OD!'OXFG!LIHADH!FLEG4CB!GFBIMHFS!WGCL!BJFMFHAM! LIBXMF!T4QCQMABOAR!GAHTFG!HTAD!AD!ODAEOMOH4!HC! QAHHFGD!BJFMFHAM!LIBXMF!CG!BQFXOW4!L4CWOEFG!H4QFZ! Defective Morphogenesis and Maintenance of Dicer Mutant Myofibers 1C!ASSGFBB!OW!'OXFG!OB!OLQCGHADH!WCG!L4CW OEFG!WCGLAHOCD!CG!LAHIGAHOCD![F!F]ALODFS! L4CWOEFG!SFNFMCQLFDH!OD!MOLE!ADS!HCDPIF!LI BXIMAHIGF!IBODP!OLLIDCTOBHCXTFLOBHG4![OHT! ADHOECSOFB!BQFXOWOX!WCG!SFBLODR!A!QAD!L4CWOEFG!LA GJFG!b:OPZ!"%_cZ!!(DOH OAHOCD!CW!L4CWOEFG! WCGLAHOCD!AH!."_Z_!OD!MOLE!b:OPZ!"%_+c!ADS!HCDP IF!b:OPZ!"%_3c!AQQFAGFS!BOLOMAG!OD!LIHADH! FLEG4CB!XCLQAGFS!HC!XCDHGCMZ!!VF!CEBFGNFS! BLAMM!XOGXIMAG!SFBLOD%QCBOHONF!L4CHIEFB![OHT! XFDHGAMO\FS!DIXMFO!HTAH!AGF!ODSOXAH ONF!CW!OLLAHIGF!WOEFGBZ!!34! ."dZ_R!FAGM4!LAHIGAHOCD![AB!LOMSM4! AWWFXHFS!OD!LIHADH!WOEFGB!OD!MOLE!ADS!HCDPIF!bS AHA!DCH!BTC[DcZ!&CDHGCM!ADS!LIHADH!WOEFGB!TAS! ODXGFABFS!OD!BO\F!ADS!HTF!LAiCGOH4!CW!DIXMFO!LOPGAH FS!HC!HTF!QFGOQTFG4Z!!9C[FNFGR!LIHADH!WOEFGB! [FGF!BMOPTHM4!SOBCGPADO\FS!XCLQAGFS!HC!XCDHGC MZ!!1TF!."dZ_!XCDHGCM!WOEFGB!EFPAD!HC!EFXCLF! CGPADO\FS!OD!GC[B![TOMF!LIHADH !WOEFGB!BTC[FS!MFBB!CGPADO\A HOCD!ADS!HTOB!XCDHODIFS!OD!."`Z_! FLEG4CB!ADS![AB!FBQFXOAMM4!DCHOXFAEMF!OD!HCDPI F!b:OPZ!"%_3cZ!!!1TF!."`Z_!XCDHGCM!HCDPIFB! SOBQMA4FS!A!GFPIMAG!MAHHOXF!CW! MCDPOHISODAM!ADS!HGADBNFGBF!L4CW OEFGB!HTAH![AB!DCH!CEBFGNFS!OD! LIHADHBZ!!:IGHTFGLCGFR!HTF!LCGQTCMCP4!CW!LIHADH! WOEFGB![AB!XCDBOSFGAEM4! SOWWFGFDH!HTAD!XCDHGCM! AH!MAHFG!SFNFMCQLFDHAM!HOLF!QCODHBZ!!,IHADH!WOEFGB! MAXJFS!SFBLOD!BHAODODP!OD!HTF!XFDHFGR!NAGOFS!OD! BO\F!ADS![FGF!OGGFPIMAGM4!BTAQFSZ!!1TFBF!GFBIMHB! ODSOXAHFS!HTAH!'OXFG!AXHONOH4![AB!SOBQFDBAEMF!WCG! ODOHOAHODP!L4CWOEFG!WCGLAHOCDR!EIH![AB! GFYIOGFS!WCG!L4CWOEFG!LCGQTCPFDFBOBZ!

PAGE 20

! =>! Increased Apoptosis in Dicer Mutant Muscle (DXGFABFS!XFMM!SFAHT!TAB!EFFD!CEBFGNFS![TFD !'OXFG![AB!ODAXHONAHFS!OD!MOLE!LFBCSFGLR! MIDP!FQOHTFMOIL!ADS!XIMHIGFS!XFMMB!b_aRd=Rd?cZ!!1T FGFWCGFR![F!T4QCHTFBO\F S!HTAH!ODXGFABFS!XFMM! SFAHT!CW!L4CPFDOX!XFMMB!XCDHGOEIHFS! HC!HTF!SFXGFABFS!BJFMFHAM!LIBXMF !OD!LIHADH!FLEG4CBZ!!1C!HFBH! HTOB!T4QCHTFBOBR![F!QFGWCGLFS!17#.-R!A!BQFXOWOX! LAGJFG!CW!'#+!WGAPLFDHAHOCD!ABBCXOAHFS![OHT! AQCQHCHOX!DIXMFOR!CD!XCDHGCM!ADS!LIHADH!WCGFMOLE!B FXHOCDB!AH!SOWWFGFDH!SFNFMCQLFDHAM!BHAPFB!b:OPZ! "%dcZ!!+H!."KZ_R!ECHT!XCDHGCM!ADS! LIHADH!MOLE!LIBXMFB![FGF!CEBFGNFS!HC!TANF!AD!AQQGFXOAEMF! DILEFG!CW!AQCQHCHOX!DIXMFO!AMHTCIPT!HTF!DILEFG![AB! BMOPTHM4!TOPTFG!OD!LIHA DH!BJFMFHAM!LIBXMF! b:OPZ!"%d+cZ!!34!."UZ_R!HTF!DILEFG!CW!17#.-!QCB OHONF!DIXMFO!SOLODOBTFS!SGABHOXAMM4!OD!XCDHGCM! FLEG4CB!EIH!GFLAODFS!TOPT!OD!LIHADHB!b0o>Z>>"cZ!! VF!CEBFGNFS!A!BOLOMAG!BOPDOWOXADH!ODXGFABF!OD! AQCQHCBOB!OD!LIHADH!BJFMFHAM! LIBXMF!XCLQAGFS!HC!XCDHGCM!AH !."_Z_!ADS!."dZ_Z!!34!."aZ_R!HTF! DILEFG!CW!AQCQHCHOX!DIXMFO![AB!BO LOMAGM4!MC[!OD!XCDHGCM!ADS!LIHADH !BJFMFHAM!LIBXMFZ!!1TOB!OB! LCBH!MOJFM4!EFXAIBF!HTF!LAiCGOH4!CW!LIHADH !LIBXMF!XFMMB!TAS!AMGFAS4!SOFSZ!!!!! +MHTCIPT!HTF!AQCQHCHOX!DIXMFO![FGF !QGFBFDH!OD!BJFMFHAM!LIBXMF!LABBFBR![F![ADHFS!HC! SFHFGLODF!OW!HTF!17#.-!QCBOHONF!DIXMFO![FGF!ODSFFS!L4CDIXMFOZ!!7DWCGHIDAHFM4R![F![FGF!IDAEMF! HC!SFHFXH!,4C'!CG!L4CPFDOD! OD!17#.-%QCBOHONF!DIXMFO!QGCEAEM 4!SIF!HC!AQCQHCBOB%ABBCXOAHFS! QGCHFOD!SFPGASAHOCD![TOXT!QGFXFS FB!'#+!WGAPLFDHAHOCDZ!+B!AD!AM HFGDAHONFR![F!F]ALODFS!HTF! MFNFM!CW!XMFANFS!XABQABF%KR!AD!FAG MOFG!LAGJFG!CW!XFMM!AQCQHCBOBZ!! 'OXFG!LIHADHB!BTC[FS!ODXGFABFS! XMFANFS!XABQABF%K!MFNFMB!XCLQAGFS!HC!XCDHGCMB! b:OPZ!"%d3R!&cZ!!(LQCGHADHM4R!HTF!LAiCGOH4!CW! XMFANFS!XABQABF%K!QCBOHONF!XFMMB! [FGF!QGFBFDH![OHTOD!BJFMFHAM !LIBXMFB!AMHTCIPT!HTF!XMFANFS! XABQABF%K!BHAODODP!SOS!DCH!AQQFAG!HC!XCLQMFHFM4 !CNFGMAQ![OHT!,9&!CD!. "_Z_!LIHADH!HGADBNFGBF! BFXHOCDB!b:OPZ!"%d&!ODBFHcZ!!9C[FNFGR!BAPOHHAM !MOLE!BFXHOCDB!GFNFAMFS! HTAH!LIHADH!L4CWOEFGB! F]QGFBBFS!ECHT!XMFANFS!XABQABF%K!ADS!,9&!A DS!HTAH!HTF!XMFANFS!XABQABF%K!BOPDAM![AB!DCH!

PAGE 21

! ="! TCLCPFDFCIBM4!SOBHGOEIHFS!HTGCIPTC IH!ODSONOSIAM!L4CWOEFGB!b:OPZ!"% d'cZ!!1TOB!GFBIMH!XCDWOGLFS! HTAH!'OXFG!LIHADH!BJFMFHAM!LIBX MF!XFMMB![FGF!ODSFFS!S4ODPZ!!! Loss of Dicer in Myoblasts also Leads to Enhanced Cell Death VF!AMBC!ODNFBHOPAHFS!HTF!FWWFXH!CW!'OXFG!OD AXHONAHOCD!CD!L4CPFDOX!SOWWFGFDHOAHOCD!OD!NOHGCZ!! VF![FGF!IDAEMF!HC!SFGONF!L4CEMABHB!WGCL!LIHADH!FLEG4CBR!AMHTCIPT![F!BIXXFBBWIMM4!XIMHIGFS! XCDHGCM!L4CEMABHBZ!!1TOB![AB!MOJFM4!SIF!HC!OLQAOG FS!PGC[HT!ADShCG!ODXGFA BFS!AQCQHCBOB!CW!LIHADH! L4CEMABHBZ!!1C!XOGXILNFDH!HTOB!QGCEMFLR![F! XCMMAECGAHFS![OHT!2AGA!)FCGPFB!WGCL!2HFQTFD! 1AQBXCHH6B!MAECGAHCG4!HC!XCDNFGH!'OXFG!XCDSO HOCDAM!LCIBF!FLEG4CDOX!WOEGCEMABHB!b,.:Bc!HC! L4CEMABHB!E4!,4CS!CNFGF]QGFBBOCD!ADS!LIHAHFS!'O XFG!E4!FXHCQOXAMM4!F]QGFBBODP!&GF!HC!4OFMS!A! GFMAHONFM4!QIGF!QCQIMAHOCD!CW!'O XFG!LIHADH!L4CEMABHBZ!!,CGF!BQFXOWOXAMM4R![F!ODWFXHFS! BQCDHADFCIBM4!OLLCGHAMO\FS!'OXFG!XCDSOHOCDAM!WO EGCEMABHB![OHT!A!GFHGCNOGAM!XCDBHGIXH!HTAH! XCDBHOHIHONFM4!F]QGFBBFB!A!WIBOCD!QGCHFOD!EFH[ FFD!LCIBF!,4CS!ADS!HTF!TCGLCDF!EODSODP!SCLAOD! CW!HTF!FBHGCPFD!GFXFQHCG!b`>cZ!!7DSFG!XCDSOHO CDB!CW!MC[!BFGIL!ADS!OD!HTF!QGFBFDXF!CW! %FBHGASOCMR! HTF!HGADBXGOQHOCDAM!AXHONOH4!CW! HTF!,4CS%.*!WIBOCD!b,'.*c!OB!ODSIXFS!ADS!OB!BIWWOXOFDH!HC! XCDNFGH!WOEGCEMABHB!HC!HTF!LIBXMF!QTFDCH4QF!b` "cZ!!1C!ODAXHONAHF!'OXF G!OD!XCDSOHOCDAM!,'.*! ,.:BR![F!ODWFXHFS!XFMMB![OHT!AD !ASFDCNOGIB!TAGECGODP! HTF!&GF!GFXCLEODABF!PFDF!GFPIMAHFS!E4!HTF! &,/!QGCLCHFG!b+S%&,/%&GFcZ!! VF!XCDWOGLFS!'OXFG!F]CD!=U!GFXCLEODAHOCD!OD!+S%&,/%&GF! XIMHIGFB!E4!PFDCLOX!0&*!ADAM4BOB! bSAHA!DCH!BTC[DcZ!!VF!AMBC!SFHFGLODFS!OW!LIBXMF!LO*#+! MFNFMB!SFXGFABFS!OD!&GF%ODWFXHFS!'OXFG!XCDSO HOCDAM!,'.*!E4!*#+!EMCHHODP!b:OPZ!"%`+cZ!!+B! QGFNOCIBM4!GFQCGHFS!b`=cR![F!CEBFGNFS!ODXGFA BFS!MFNFMB!CW!LIBXMF%BQFXOWOX!LO*%=>d!OD! %FBHGASOCM! ODSIXFS!'OXFG!XCDSOHOCDAM!,'.*!HTAH!TANF!,4 CS!AXHONAHFSZ!!(LQCGHADHM4R!LO*%=>d!MFNFMB! SFXGFABFS![TFD!HTFBF!XFMMB![FGF !ODWFXHFS![OHT!&GFZ!!LO*%"dR! [TOXT!OB!DCH!BFDBOHONF!HC!,4CS!

PAGE 22

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b:OPZ!"%`3R!QADFMB!=!ADS!UcZ!!(LQCGHA DHM4R!HTOB!ODXGFABFS!XFM M!SFAHT![AB!SFQFDSFDH!CD! HTF!ODSIXHOCD!CW!,' .*!AXHONOH4R!AB!&GF%ODW FXHFS!XFMMB!HTAH![FGF! XIMHIGFS!OD!',![OHTCIH! % FBHGASOCM!SOS!DCH!F]TOEOH!HTF!BALF!MFNFM!CW!XFMM!SFAH T!b:OPZ!"%`3R!QADFMB!K!ADS!UcZ!!1C!YIADHOW4!HTOB! AQQAGFDH!ODXGFABF!OD!XFMM!SFAHT! ADS!HC!SFHFGLODF![TFHTFG!OH!GF WMFXHFS!ODXGFABFS!AQCQHCBOB!OD! SOWWFGFDHOAHODP!&GF%ODWFXHFS!'OXFG!XCDSOHOCDAM!,'.*!XIMHIGFBR![F!QFGWCGLFS!+DDF]OD!/% :(1&hQGCQOSOIL!OCSOSF!BHAODODP!CW!HTF!XIMHIGF B!ADS!YIADHOWOFS!HTF!QCBO HONFM4%BHAODFS!XFMMB!E4! :+&2!ADAM4BOB!b:OPZ!"%`cZ!!+DDF]OD/%:(1&!BHAO DODP!OSFDHOWOFB!AQCQHCHOX! XFMMB!E4!BQFXOWOXAMM4! SFHFXHODP!HTF!HGADBMCXAHOCD!CW!QTCBQTAHOS4MBFGODF!WG CL!HTF!ODDFG!WAXF!CW!HTF!QMABLA!LFLEGADF!HC! HTF!XFMM!BIGWAXF!HTAH!CXXIGB!SIGODP!HTF!FAGM4!BHAP FB!CW!AQCQHCBOBZ!!1C!QFGLOH!YIADHOHAHONF!XFMM! BCGHODP!E4!:+&2R![F!XTCBF!HC!F]ALODF!AD!FAGM4 !BHAPF!CW!L4CPFDOX!SOWWFGFDHOAHOCD!b=U!TCIGBc! [TFD![F!F]QFXHFS!HTAH!AQCQHCBOB!LA4!EF!ODOHOA HFSR!EIH!EFWCGF!HTF!XFMMB!TAS!WIBFS!HC!EFXCLF! LIMHODIXMFAHFS!L4CHIEFBZ!!+MM!BALQMFB![FGF!ADAM4 \FS!OD!HTF!QGFBFDXF!CW!ECHT!QGCQOSOIL!OCSOSF! ADS!+DDF]OD!/%:(1&R!ADS!XFMM!BCG HODP!GFNFAMFS!HTF!QFGXFDHAPF! CW!XFMMB!HTAH![FGF!NOAEMFR! DCDNOAEMF!bFOHTFG!AQCQHCHOX!CG!DFXGCHOXg!QGCQOSOI L!OCSOSF!QCBOHONFcR!CG!AQCQHCHOX!b+DDF]OD!/% :(1&!QCBOHONFc!b:OPZ!"%`&cZ!!&CDBOBHFDH![OHT!HT F!ODXGFABFS!XFMM!SFAHT!CEBFGNFS!OD!&GF%ODWFXHFS! 'OXFG!XCDSOHOCDAM!,'.*!XFMMB! OD!HTF!QGFBFDXF!CW!',!QMIB! %FBHGASOCMR![F!CEBFGNFS!A!PGFAHFG! HTAD!H[C%WCMS!ODXGFABF!OD!+DDF]OD!/%QCBOHONFR!AQ CQHCHOX!XFMMB![TFD!,4CS!AXHONOH4![AB!ODSIXFS!OD!

PAGE 23

! =K! HTF!AEBFDXF!CW!'OXFG!WIDXHOCDR!XCLQAGFS!HC!ID ODSIXFS!&GF%ODWFXHFS! 'OXFG!XCDSOHOCDAM!,'.*! XFMMB!b:OPZ!"%`&c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b:OPZ!"%`3R!QADFM!UcZ! .]ALODAHOCD!CW!HTF!QGCQOSOIL!OCSOSF%QCBOHONF!QCQIMAHOCD!ADS!HTF!SIAMM4!QCBOHONF!+DDF]OD! /%:(1&!ADS!QGCQOSOIL!OCSOSF!QCBOHONF!XFMMBR!GFN FAMFS!AD!AQQGC]OLAHF!"Z`%WCMS!ODXGFABF!OD!XFMM! SFAHT!OD!HTF!'OXFG %h% !,'.*!XFMMB!AH!=U!TCIGB!WCMMC[ODP!, 4CS!ODSIXHOCD!b:OPZ!"%`&cZ!!VF!XADDCH! AHHGOEIHF!HTFBF!XTADPFB!SOGFXHM4!HC!AQCQHCBOBR!EIH! EFXAIBF!QGCQOSOIL!OCSOSF!XAD!FDHFG!XFMMB!OD!HTF! MAHFG!BHAPFB!CW!AQCQHCBOBR!OH!OB! MOJFM4!HTAH!HTF!SCIEM4!QCBOHONF!XF MMB!GFQGFBFDH!MA HF!BHAPFB!CW! AQCQHCBOB!GAHTFG!HTAD!DFXGCBOBZ!!+B!HTOB!FWWFXH ![AB!DCH!CEBFGNFS!OD!'OXFG !XCDSOHOCDAM!,'.*!XFMMB! WCMMC[ODP!HTF!ODSIXHOCD!CW!,4CS!AXHONOH4R!ADS!AB! HTF!QFGXFDHAPF!CW!QGCQOSOIL!OCSOSF%QCBOHONF! XFMMB![AB!XCLQAGAEMF!OD!ECHT!'O XFG!XCDSOHCDAM!,'.*!ADS!&GF%ODWF XHFS!'OXFG!XCDSOHOCDAM!,'.*! XFMMB!OD!HTF!AEBFDXF!CW!,4CS!AXHONOH4R![F!AHHGOEIH F!HTOB!FWWFXH!BQFXOWOXAMM4 !HC!,4CS!AXHONAHOCD!OD! HTF!AEBFDXF!CW!'OXFG!WIDXHOCDZ!!!!! $NFGAMMR!HTFBF!GFBIMHB!AGF!XCDBOBHFDH![OHT!ECH T!HTF!ODXGFABFS!XFMM!SFAHT!HTAH![F!CEBFGNFS!OD! MAHF%BHAPF!SOWWFGFDHOAHFS!&GF%ODW FXHFS!'OXFG!XCDSOHOCDAM!,'.*!XIMHI GFB!OD!NOHGC!b:OPZ!"%`3R!QADFM! Uc!ADS![OHT!HTF!ODXGFABFS!AQCQHCBOB !BFFD!OD!'OXFG!LIHADH!LIBXMF!OD !NONCZ!!1TFGFWCGFR ![F!XCDXMISF! HTAH!HTF!MCBB!CW!'OXFG!WIDXHOCD!OD !BJFMFHAM!LIBXMF!QGCLCHFS!ODXG FABFS!AQCQHCBOB!OD!ECHT!L4CWOEFGB! ADS!L4CEMABHBZ!

PAGE 24

! =U! Decreased Cell Proliferation Foll owing Loss of Muscle MiRNAs 0GFLAHIGF!SOWWFGFDHOAHOCD!CW!NFDHGOXIMAG!XA GSOCL4CX4HFB![AB!CEBFGNFS![TFD!A!LO*%"! HGADBPFDF![AB!CNFGF]QGFBBFS!OD!LCIB F!TFAGH!HOBBIF!b`KcZ!!1TFGFWCG FR![F!QCBHIMAHFS!HTAH!GFSIXFS! MFNFMB!CW!LO*%"!CEBFGNFS!OD!CIG!'OXFG!BJFMFHAM!LIBX MF!LIHADHB!GFBIMHFS!OD!HT F!ODAEOMOH4!CW!BJFMFHAM! LIBXMF!QGFXIGBCGB!HC!SOWWFGFDHOAHFR![TOXT!IMHOLAH FM4!MFS!HC!MFBB!LIBXMF!CNFGAMMZ!!VF!MAEFMFS! QGCMOWFGAHODP!XFMMB!OD!XCDHGCM!ADS!LIHADH! MOLE!LIBXMFB![OHT!AD!ADHOECS4!BQFXOWOX!WCG! QTCBQTCG4MAHFS!TOBHCDF!9K!IBODP! XCDHGCM!ADS!LIHADH!MOLE!BFX HOCDB!HTGCIPTCIH!FLEG4CPFDFBOB! b:OP!"%acZ!!1TF!."KZ_!XCDHGCM!ADS!LIHADH!MOLEB!TA S!A!BOLOMAG!DILEFG!CW!QG CMOWFGAHODP!XFMMB!b:OPZ! "%a+cZ!!1TF!DILEFG!CW!QGCMOWFG AHODP!XFMMB!ODXGFABFS!OD!XCDHGCMB !WGCL!."UZ_%."dZ_!b:OPZ!"%a3cZ!! 9C[FNFGR!XFMM!QGCMOWFGAHOCD!S FXGFABFS!OD!LIHADH!LIBXMF!SIGODP!HTOB!HOLFWGALF!ADS![AB! BOPDOWOXADHM4!GFSIXFS!XCLQAGFS!HC !XCDHGCMB!b0o>Z>>"cZ!!+H!."`Z_R! HTF!DILEFG!CW!QGCMOWFGAHODP! XFMMB!SFXGFABFS!OD!XCDHGCM!FLEG4CB!ADS![AB!BMOPT HM4!MFBB!OD!LIHADHB!XCLQAGFS!HC!XCDHGCMBZ!!34! ."aZ_R!ECHT!XCDHGCM!ADS!LIHADH!BJFM FHAM!LIBXMF!TAS!BOLOMAGM4!MC[! MFNFMB!CW!XFMM!QGCMOWFGAHOCDZ!! 1TFGFWCGFR!ODAXHONAHOCD!CW!'OXFG!MFAS!HC!SFXG FABFS!QGCMOWFGAHOCD!CW! BJFMFHAM!LIBXMF!XFMMBZ! Myogenic Regulatory Factors are Aberrantly Ex pressed after Dicer Inactivation !VF!F]ALODFS!HTF!F]QGFBBOCD!CW!BFNFGAM!WAXHCGB! HTAH!AGF!OLQCGHADH!SIGO DP!SOWWFGFDH!BHAPFB!CW! L4CPFDFBOB!OD!AD!AHHFLQH!HC!SF MODFAHF!HTF!LCMFXIMAG!SFWFXHB!ABBCXOAHFS![OHT!AEDCGLAM!LIBXMF! LCGQTCPFDFBOBZ!!(LLIDCEMCHHODP![AB!QFGWCGLFS! CD!."aZ_!XCDHGCM!ADS!LIHADH!HCDPIF!F]HGAXHB! b:OP!"%?cZ!!,4C'!ADS!L4CPFDOD![FGF!XTCBFD!EFXAI BF!HTF4!AGF!HGADBXGOQHO CD!WAXHCGB!HTAH!ODOHOAHF! HTF!L4CPFDOX!XABXASFZ!!,4CBOD!TFAN4!XTAOD!b,9&c! ADS!SFBLOD![FGF!AMBC!ADAM4\FS!EFXAIBF!HTF4! AGF!L4CWOEFG!BHGIXHIGAM!XCLQCDFDHB!HTAH!AGF! SC[DBHGFAL!OD!HTF!L4CPFDOX!QGCPGALZ!!+H!."aZ_R! ,4C'!ADS!L4CPFDOD!QGCHFOD!MFNFMB! [FGF!TOPTFG!OD!LIHADH!BJFMFHAM!LIBXMF!XCLQAGFS!HC!XCDHGCMZ!! (DHFGFBHODPM4R!,9&![AB!SC[DGFPIMAHFS!OD!LI HADH!FLEG4CB![TOMF!SFBLOD!F]QGFBBOCD![AB! IDAWWFXHFSZ!

PAGE 25

! =_! Discussion 'OXFG!ADS!LO*#+B!AGF!OLQCGHADH!WCG!A!NAGOFH 4!CW!SFNFMCQLFDHAM!QGCXFBBFB!b`U%`dcZ!!1TF! CEiFXHONF!CW!HTOB!BHIS4![AB!HC!SFHFGLODF!HTF!GCMF !CW!'OXFG!ADS!LO*#+%LF SOAHFS!PFDF!GFPIMAHOCD! OD!BJFMFHAM!LIBXMFZ!!$IG!ADAM4BOB!SFLCDBHGAHFB! HTAH!'OXFG!AXHONOH4!OB!FBBFDHOAM!WCG!QCBH%DAHAM!MOWF! ADS!BJFMFHAM!LIBXMF!SFNFMCQLFD H!SIGODP!FLEG4CPFDFBOBZ!!'OXFG!OD AXHONAHOCD!OD!BJFMFHAM!LIBXMF! GFBIMHB!OD!MC[FG!MFNFMB!CW!LIBXMF%BQFXOWOX!LO*# +B!ADS!'OXFG!LIBXMF!LIHADHB!SOF!QFGODAHAMM4!ADS! AGF!XTAGAXHFGO\FS!E4!BJFMFHAM!LI BXMF!T4QCQMABOAZ!!*FSIXFS!BJFMFH AM!LIBXMF!OB!AXXCLQADOFS!E4! AEDCGLAM!L4CWOEFG!LCGQTCMCP4Z!!1TF!BJFMFHAM!LIBX MF!SFWFXHB!ABBCXOAHFS![OHT!MCBB!CW!'OXFG! WIDXHOCD!LA4!EF!F]QMAODFS!E4!ODXGFABFS!AQCQHCB OB!HCPFHTFG![OHT!SFXGFABFS!DILEFGB!CW! QGCMOWFGAHODP!LIBXMF!XFMMBZ!!1TOB !BHIS4!OB!HTF!WOGBH!HC!QGCNOSF! 57'858" !FNOSFDXF!CW!HT F!OLQCGHADXF!CW! 'OXFG!WIDXHOCD!ADS!LO*#+%LFSOAHFS!PFDF!BO MFDXODP!SIGODP!LALLAMOAD!BJFMFHAM!LIBXMF! SFNFMCQLFDHZ! Dicer is Essential for Skeletal Muscle Morphogenesis 1TF!QTFDCH4QF!ABBCXOAHFS![OHT!'OXFG!ODAXHONAH OCD!ADS!GFSIXFS!LIBXMF!LO*#+B!OD!LOXF!OB! BOLOMAG!HC!HTAH!CEBFGNFS!OD! +&"E"?$5=)',5IF!b J,5&Fc!LIHADHB!b``R`acZ!!8DCXJCIH! J,5&F-' WMOFB! AGGFBH!SIGODP!FLEG4CPFDFBOB!CG! MAGNAM!SFNFMCQLFDHR!SOBQMA4!SOBCGPADO\FS!LIBXMF!LCGQTCMCP4! ADS!TANF!AEFGGADH!F]QGFBBOCD!CW!LIBXMF%BQFXOWO X!PFDFBZ!!:IGHTFGR!SLOG%"!HAGPFHB!'FMHAR!A! LFLEGADF%ECIDS!MOPADS!WCG!#CHXT!ADS! J,5&F-' LIHADHB!TANF!SOBGIQHFS!'FMHA%#CHXT!BOPDAMODPZ!! #CHXT!BOPDAMODP!AMBC!QMA4B!AD!OLQCGHADH!GCMF !OD!LALLAMOAD!L4CPFDFBOB!b`?cZ!!1TFGFWCGFR! QCHFDHOAM!SFWFXHB!OD!#CHXT!BOPDAMODP!OD!LCIBF! +56%&' LIHADH!FLEG4CB!LA4!XCDHGOEIHF!HC!AEDCGLAM! LIBXMF!LCGQTCPFDFBOBZ!!1TFBF!CEBFGNAHOCDB!AGPI F!HTAH!LO*%"!QMA4B!AD!FBBFDHOAM!GCMF!OD! FLEG4CDOX!LIBXMF!SFNFMCQLFDHZ! (DHFGFBHODPM4R! +56%&' BJFMFHAM!LIBXMF!LIHADHB!AGF!CEBFGNFS !HC!TANF!GFSIXFS!BJFMFHAM!LIBXMF! OD!HTF!QGFBFDXF!CW!ODXGFABFS!MFNFMB!CW! ,*:BZ!!,4C'R!L4CPFDOD!ADS!,FW=!QGCLCHF! ,5IFR! ,5IF

PAGE 26

! =d! -KK !ADS! ,5IFLMN' F]QGFBBOCD!E4!EODSODP!HC!IQBHGFAL! 65E !FMFLFDHB!ba>cZ!!1TOB!GAOBFB!HTF!QCBBOEOMOH4! HTAH!,*:B!AGF!IQGFPIMAHFS!HC!XCLQFDBAHF!WCG!GFSIXFS!LO*#+!MFNFMB!OD! +56%&' LIHADHBZ!! 9C[FNFGR!HTOB!SCFB!DCH!F]QMAOD![T4!ODXGFAB FS!,*:B!WAOM!HC!QGCLCHF!L4CPFDFBOB!OD! +56%&' LIHADH! BJFMFHAM!LIBXMFZ!!$DF!QCBBOEOMOH4 !OB!HTAH!L4CPFDOX!ODTOEOHCG4!W AXHCGB!HTAH!ADHAPCDO\F!,*:B!AGF! IQGFPIMAHFS!OD! +56%&' LIHADHBZ!!.]ALQMFB!CW!BIXT!WAXHCG B!AGF!,4C*!ADS!(S!WALOM4!LFLEFGB! ba"Ra=cZ!!1TFBF!QGCHFODB!WIDXHO CD!AB!L4CPFDOX!ADHAPCDOBHB!E4!O DTOEOHODP!EODSODP!CW!,*:B!HC!.! EC]FB!OD!HTF!QGCLCHFGB!CW!LIBXMF%BQFXOWOX!PFDFBZ !!(DSFFSR!CNFGF]QGFBBOCD!CW!LO*%=>d!OD!XIMHIGFS! L4CEMABHB!GFBIMHB!OD!SF XGFABFS!F]QGFBBOCD!CW! !O"I R! 2JR! 2JL !ADS! 2JK' baKcZ!!1TIBR!LIBXMF! LO*#+B!LA4!QGCLCHF!L4CPFDFBOB!E4!SC[DGFPIMA HODP!F]QGFBBOCD!CW!ODTOEOHCG4!WAXHCGBZ!! 1TF!SFXGFABF!OD!LIBXMF!LABB!OD! +56%&' LIHADHB!OB!BHGOJODPM4!BOLOMAG!HC!QTFDCH4QFB! ABBCXOAHFS![OHT!LIBXIMAG!S4BHGCQTOFB!ADS!APFS!BJFM FHAM!LIBXMFZ!!(H![OMM!EF !ODHFGFBHODP!HC!BFF!OW! 'OXFG!LIHAHOCDBR!CG!SOBGIQHFS!LO *#+%LFSOAHFS!PFDF!GFPIMAHOCDR!X CDHGOEIHF!HC!BJ FMFHAM!LIBXMF! L4CQAHT4!ADS!APF%GFMAHFS!BAGXCQFDOAZ!! Dicer is Required for Muscle Cell Survival and Proliferation during Embryogenesis (DXGFABFS!AQCQHCBOB!OD! +56%& !LIHADH!BJFMFHAM!LIBXMF!QGCNOS FB!CDF!F]QMADAHOCD![T4!LIHADH! FLEG4CB!AGF!XTAGAXHFGO\FS!E4!BJFMFHAM!LIBXMF !T4QCQMABOA!ADS!AEDCGLAM!L4CWOEFG!LCGQTCMCP4Z!! 9C[FNFGR!HTF!GFMAHOCDBTOQ!EFH[FFD!'OXFGR!LO*#+B !ADS!AQCQHCBOB!OB!IDXMFAGZ!!0GCPGALLFS!XFMM! SFAHT!CXXIGB!OD!A!NAGOFH4!CW! XFMM!H4QFB!SIGODP!FLEG4CPFDFBOB!ADS! OB!HTCIPTH!HC!EF!GFYIOGFS!WCG! GAQOS!HOBBIF!GFLCSFMODP!baUcZ!!0G CSIXHONF!HOBBIF!PGC[HT! SIGODP!SFNFMCQLFDH!GFYIOGFB!HTF!GAHF!CW! XFMM!QGCMOWFGAHOCD!HC!F]XF FS!HTAH!CW!XFMM!SFAHT!ADS!LO*#+%LF SOAHFS!PFDF!BOMFDXODP!LA4!WIDXHOCD! HC!GFBHGOXH!CG!ODTOEOH!AQCQHCBO B!SIGODP!FLEG4CDOX!SFNFMCQLFDH!E4!HAGP FHODP!QGC%AQCQHCHOX!WAXHCGBZ!! LO*%="!TAB!ADHO%AQCQHCHOX!AXHONOH4!OD!PMOCEMAB HCLA!XFMMB!ADS!BFNFGAM!CHTFG!LO*#+B!AGF! OLQMOXAHFS!OD!HILCG!BIQQGFBBOCD!ba_cbadcZ!!(H!OB !QCBBOEMF!HTAH!LIBXMF! LO*#+B!TANF!AD!ADHO% AQCQHCHOX!AXHONOH4!HTAH!OB!GFYIOGFS!HC!FDBIGF!HT AH!HTF!AQCQHCBOB!GFYIOGFS! AH!FAGM4!SFNFMCQLFDHAM!

PAGE 27

! =`! HOLF!QCODHB!SCFB!DCH!QFGBOBH!SIGODP!MA HFG!QFGOCSB!CW!GAQOS!QGCMOWFGAH OCD!ADS!PGC[HTZ!!:CG!F]ALQMFR! LIBXMF!LO*#+bBc!LA4!WIDXHOCD!OD!HTF!SFXMODF!CW! AQCQHCBOB!CEBFGNFS!OD! MOLE!BJFMFHAM!LIBXMF! EFH[FFD!."KZU!ADS!."UZ_!b:OP!"%dc!E4!ODTOEOHODP!HTF! F]QGFBBOCD!CW!QGC%AQCQHCHOX!WAXHCGBZ!!-CBB!CW! HTOB!GFPIMAHOCD!LA4!HTFD!GFBIMH!OD!ODXGFABFS! AQCQHCBOB!ADS!SFXGFABFS!BJFMFHAM!LIBXMFZ!!!! +MHFGDAHONFM4R!ODXGFABFS!AQCQHCBOB!OD! +56%&' LIHADH!LIBXMF!LA4!EF!ODSFQFDSFDH!CW!LO*#+B! ADS!GFBIMH!WGCL!IDXTAGAXHFGO\FS!'O XFG!WIDXHOCDBZ!!-CBB!CW!'OXFG!OD !MOLE!ADS!MIDP!AMBC!GFBIMHB!OD! ODXGFABFS!XFMM!SFAHT!ADS!LCGQTCMCPOXAM!SFWFXHB! b_aRd=cZ!!(D!WOBBOCD!4FABHR!'OXFG!ADS!*#+! ODHFGWFGFDXF!AGF!OLQCGHADH!WCG! GFPIMAHODP!XTGCLCBCLF!BFPGFPAHOCD!ADS!TFHFGCXTGCLAHOX!PFDF! BOMFDXODP!ba`RaacZ!!1TFGFWCGFR!'O XFG!ODAXHONAHOCD!LOPTH!MFAS!HC! QFGHIGEAHOCDB!OD!XTGCLCBCLF! BFPGFPAHOCD!ADS!XCDSFDBAHOCD![TOX T!HGOPPFG!XFMM!SFAHT!HC!ANCOS!AE FGGADH!XFMM!SONOBOCD!ADS!PFDF! F]QGFBBOCDZ! *FSIXFS!QGCMOWFGAHONF!QCHFDHOAM!CW!BJFMFHAM !LIBXMF!QGFXIGBCGB!LA4!AMBC!F]QMAOD![T4! +56%& LIHADHB!QCBBFBB!MFBB!LIBXMFZ!!'FXGFABFS!DILEFGB! CW!QGCMOWFGAHODP!XFMMB!OD!LIHADH!LIBXMF! HTGCIPTCIH!FLEG4CPFDFBOB!XAD!EF!F] QMAODFS!E4!ODXGFABFS!XFMM!SFAHT! CW!XFMMB!HTAH![CIMS!DCGLAMM4! EF!AXHONFM4!SONOSODPZ!!+D!AMHFGDAH ONF!F]QMADAHOCD!OB!HTAH!LO*#+B! GFPIMAHF!XFMM!QGCMOWFGAHOCD!ADShCG! SOWWFGFDHOAHOCDR!ADS!SOBGIQHOCD!CW!LO*#+!QGCXFBBO DP!GFBIMHB!OD!SFXGFABFS! XFMM!QGCMOWFGAHOCD!OD! +56%& !LIHADH!BJFMFHAM!LIBXMFZ!!.NOSFDXF!BIQQCGHODP! HTOB!MAHHFG!QCBBOEOMOH4!XCLFB!WGCL!BHISOFB! IBODP!&=&"=!L4CEMABHB!ADS! 3%7"?*E !FLEG4CB!badcZ!!LO*%"!ADS! LO*%"KK!TANF!ADHAPCDOBHOX! FWWFXHB!OD!XIMHIGFS!XFMMBZ!!LO*%"!FDTADXFB! L4CPFDOX!SOWWFGFDHOAHOCD![TOMF!LO*%"KK!QGCLCHFB! L4CEMABH!QGCMOWFGAHOCDZ!!1T FGFWCGFR!HTF!SFXGFABF!OD!QGCMOWFGA HODP!XFMM!DILEFG!ADS!BJFMFHAM!LIBXMF! T4QCQMABOA!OD! +56%& !LIHADHB!LA4!EF!AHHGOEIHFS!HC!GFSIXFS !MFNFMB!CW!WIDXHOCDAM!LO*%"KK!ADS!LO*% "R!GFBQFXHONFM4Z!!(H!OB!AMBC!ODHFGFBHODP!HC!DCH F!HTAH!LO*%"!ADS!LO*%=>d!TANF!OSFDHOXAM!BFFS! BFYIFDXFB!HTAH!GFXCPDO\F!L*#+!K6 71*!HAGPFHB!HC!ODTOEOH!QGCHFOD! F]QGFBBOCD![TOXT!BIPPFBHB!HTAH!

PAGE 28

! =a! LO*%"!ADS!LO*%=>d!BTAGF!BCLF!SFPGFF!CW!WIDXHO CDAM!GFSIDSADX4Z!!+SSOHOCDAMM4R!LO*%=>d!HAGPFHB! HTF!MAGPF!BIEIDOH!CW!'#+!QCM4LFGABF!b /"=)c!OD!XIMHIGFS!XFMMB!baKcZ!!1TF!QCHFDHOAM!WCG!ODXGFABFS! F]QGFBBOCD!CW! /"=)-' SIF!HC!GFSIXFS!MFNFMB!CW!LO*%=>d!OD! +56%&' LIHADHB!LA4!GFBIMH!OD! IDXCDHGCMMFS!'#+!GFQMOXAHOCD!ADS!XFMM!SFAHTZ!!&ILIMA HONFM4R!HTFBF!SAHA!SFLCDB HGAHF!HTAH!BJFMFHAM! LIBXMF!LO*#+B!LA4!GFPIMAHF!HTF!EAMADXF!EFH[ FFD!L4CPFDOX!QGCMOWFGAHOCDR!SOWWFGFDHOAHOCD!ADS! XFMM!SFAHT!SIGODP!FLEG4CDOX!L4CPFDFBOBZ!!(DHFGF BHODPM4R!BFNFGAM!LO*#+B !ADS!LO*#+!XMIBHFGB! AGF!ABBCXOAHFS![OHT!BQFXOWOX!XADXFGB!ba?cZ!!! 'OXFG6B!LO*#+!QGCXFBBODP!AEOMOH4!TAB!EFFD!B TC[D!HC!EF!FBBFDHOAM!WCG!MOLER!MIDPR!BJOD!ADS! TAOG!SFNFMCQLFDHZ!!$IG!GFBIMHB!SFLCDBHGAHF!HTAH !'OXFG!ADS!LO*#+B!AGF!AMBC!ODSOBQFDBAEMF!WCG! FLEG4CDOX!BJFMFHAM!LIBXMF!SFNFMCQLFDHZ!!(H![ OMM!EF!ODHFGFBHODP!HC!B FF!OW!'OXFG!ADS!LO*#+B! WIDXHOCD!SIGODP!QCBH%DAHAM!L4CPFDFBOB!ADS!L4CWOEFG!GFPFDFGAHOCDZ!!!!!

PAGE 29

! =?! 1AEMF!"%"Z!!LO*#+B!OSFDHOWOFS!OD! A!NAGOFH4!CW!SOWWFGFDH!CGPADOBLB! 2QFXOFB!&CLLCD!DALF!#ILEFG!CW!LO*#+B +DCQTFMFB!PALEOAF!,CBYIOHC!_=! 'GCBCQTOMA!LFMADCPABHFG!:M4!"_=! &AFDCGTAESOHOB!FMFPADB! eFDCQIB!MAFNOB! VCGL! :GCP! "_U! `! eFDCQIB!HGCQOXAMOB!:GCP!"aU! )AMMIB!PAMMIB!&TOXJFD!U`_! &ADOB!WALOMOAGOB!'CP!=>U! ,AXAXA!LIMAHHA!,CDJF4!UdK! )CGOMMA!PCGOMMA!)CGOMMA!ad! 9CLC!BAQOFDB!9ILAD!d?_! 0AD!HGCPMCS4HFB!&TOLQAD\FF!_?_! ,IB!LIBXIMIB!,CIBF!Uaa! *AHHIB!DCGNFPOXIB!*AH!=ad! 3CB!HAIGIB!&C[!""`! 'ADOC!GFGOC!mFEGAWOBT!KK`! :IPI!GIEGOQFB!0IWWFGWOBT!"K"! +GAEOSCQBOB!HTAMOADA! .QBHFOD!3AGG!NOGIB! 9FGQFB!2OLQMF]!NOGIB! 9ILAD!X4HCLFPAMCNOGIB! :MC[FGODP!QMADH! /OGIB! /OGIB! /OGIB! "a`! =K! d! K! !1TOB!SAHA!OB!WGCL!LO*3ABF!CD!5ADIAG4!"_R!=>>?Z! THHQfhhLOXGCGDAZBADPFGZAXZIJhXPO%EODhBFYIFDXFBhEGC[BFZQM

PAGE 30

! K>! :OPIGF!"%"Z!LO*#+!EOCPFDFBOBR!LAHIGAHOCD!ADS!HGADBMA HOCD!ODTOEOHOCDZ!!LO*#+B!AGF!HGADBXGOEFS!!!!! E4!*#+!QCM4LFGABF!((!ODHC!LCDCXOBHGCDOX!CG!QCM4XOBHGCDOX!QGOLAG4!bQGOc%LO*#+B! HTAH!WCGL!SCIEMF%BHGADSFS!TA OGQOD!BHGIXHIGFB!XCDHAODODP! A!_6!`%LFHT4M!PIADCBODF!XAQ! ADS!A!K6!QCM4b+c!HAOMZ!!0GO%LO*#+B!AG F!QGCXFBBFS!OD!HTF!DIXMFIB!E4!HTF!SCIEMF% BHGADSFS!FDSCGOECDIXMFABF!'GCBTA!ODHC!k `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b*(2&cZ!!1TF!*(2&!OSFDHOWOFB! HAGPFH!L*#+B!EABFS!CD!BFYIFDXF!XCLQMFLFDHAGOH4!EFH[FFD!HTF!LO*#+!ADS!OHB! HAGPFHZ!!1TF!LO*#+!HAGPFHFS!L*#+B!AGF!HT FD!HGAWWOXJFS!HC!)V!ECSOFB![TFD!HTF4! IDSFGPC!SFPGASAHOCD!CG!HGADBM AHOCDAM!ODTOEOHOCDZ!!'FPGASAH OCD!CW!HTF!L*#+!CXXIGB!E4! HTF!BMOXFG!AXHONOH4!CW!+GPCDA IHF=R!CDF!CW!HTF!*(2&%ABBCXOAHFS!WAXHCGBZ!!+GPCDAIHF=! XMFANFB!L*#+B!AH!A!BOHF!DFAG!HT F!LOSSMF!CW!HTF!LO*#+fL*#+!T4EGOSZ!! LO*#+Bh*(&2!ODTOEOH!HGADBMAHOCD!E4!QGFNFD HODP!HTF!ODHFGAXHOCD! EFH[FFD!HTF!QCM4b+c! HAOM!ADS!HTF!_6XAQZ!!*FQGODHFS!WGCL! !5I(PE'57'+%8%="?,%7#')7J'Q*,"&5D%7%E5E g! $6*CIGJF!5*R!2[ADBCD!,2!ADS!9AGWF! 3'g!&CQ4GOPTH!=>>_Z!!*FQGODHFS![OHT! QFGLOBBOCD!CW!VOMF4%-OBBR!(DXZ!A!BIEB OSOAG4!CW!5CTD!VOMF4!n!2CDBR!(DXZp!

PAGE 31

! K"!

PAGE 32

! K=! :OPIGF!"%=Z!(DAXHONAHOCD!CW!BJFMFH AM!LIBXMF!'OXFG!ADS!SFXGFABFS! LIBXMF%BQFXOWOX!LO*#+BZ!!+c! +56%&' XCDSOHOCDAM!AMMFMF!ADS! !O"+6&%' HGADBPFDFZ!! +56%&' XCDSOHOCDAM!AMMFMF!XCDHAODB! ="A/' BOHFB!bHGOADPMFBc!WMADJODP!HTF!*#+BF! (((!SCLAOD!FDXCSFS!E4!F]CD!bWOMMFS!EC]FBc! =U!bPGA4!EC]cZ!!+!DFCL4XOD!GFBOBHADXF!XA BBFHHF!bDFCc!OB!GFHA ODFS!OD!ODHGCD!=UZ!! !O"+6&%' HGADBPFDF!XCDHAODB! B&%' GFXCLEODABF!bWOMMFS!EC]c!SC[DBHGFAL!CW!HTF! !O"+' XCGF!FDTADXFG!ADS!QGC]OLAM!QGCLCHFG! FMFLFDHB!bCQFD!EC]cZ!!3c!*1%0&*!CD!."`Z_! HCDPIF!X'#+!IBODP!QGOLFGB!WMADJODP!HAGPFH FS!F]CD!=U!bWOMMFS!EC]cZ!!0GOLFGB!bAGGC[Bc! [FGF!QCBOHOCDFS!OD!F]CD!=K!ADS!=_Z!! ,OXF!XCDHAODFS!A!XCLEODAHOCD!CW! +56%&' XCDSOHOCDAM!bXcR![OMS!H4QF!blc!CG! +56%&' SFMFHFS!F]CD!=U!b%c!AMMFMFB!ADS!A! !O"+6&%' HGADBPFDFZ!!1GADBXGOQHB!X CDHAODODP!bl.=Uc!CG!LOBBODP!F ]CD!=U!b%.=Uc!GFBIMH!OD!UK"!ADS! "??!EQ!WGAPLFDHBR!GFBQFXHONFM4Z!!&c!!*#+! EMCH!ADAM4BOB!CW!."KZ_!ADS!."UZ_!XCDHGCM! ADS!LIHADH!MOHHFGLAHF!MOLEB!HC!SFHFXH!LIBXMF%BQFXOWOX!LO*#+B!bLO*cZ!!7d!BD*#+!OB! HTF!MCASODP!XCDHGCMZ!!'OGFXH !XCLQAGOBCD!EFH[FFD!LO*!MFNFMB!XADDCH!EF!LASF!EFXAIBF! QGCEF!BQFXOWOX!AXHONOHOFB!ADS!W OML!F]QCBIGF!HOLFB!NAGOFS!WCG!FAXT!LO*Z!!*FQGODHFS!WGCL! REE%7#5)='I"=%'S"&'+56%&'J*&57D'1@%=%#)='!*E6=%'+%8%="?,%7# g!$6*CIGJF!5*R!)FCGPFB! 2+R!2FA4!9*R!1AQBXCHH!25R!,X,ADIB!,1R! )CMSTALFG!'5R!!2[ADBCD!,2!ADS!9AGWF! 3'g!&CQ4GOPTH!=>>_R![OHT!QFGLOBBOCD!WGCL!.MBFNOFGZ!

PAGE 33

! KK! ! ! :OPIGF!"%KZ! +56%&' LIHADHB!SOBQMA4!MFBB!BJFMFHAM!LIBXMFZ!! +c!2JODDFS!."aZ_!X CDHGCM!ADS!LIHADH!! MOHHFGLAHFBZ!!#CHF!HTF!OLQAOGFS!SFNFMCQLFDH!CW!LIHADH!WCGFMOLE!LIBXMFZ!!3c!e%PAM! BHAODFS!0>!XCDHGCM!ADS!LIHADH!MOHHFGLAHF! WCGFMOLEB!bHCQ!QADFMBcR!TODSMOLEB!bLOSSMF! QADFMBc!ADS!."a!HCDPIFB!bECHHCL! QADFMBcZ!!,OXF!XCDHAODFS!HTF! ILN' GFQCGHFG!OD![TOXT! e%PAM!BQFXOWOXAMM4!BHAODB!XFMMB!HTAH!F]QGFBB! !O"+6&% Z!!&c!9n.!BHAODODP!CW!XCDHGCM!ADS! LIHADH!HGADBNFGBF!SOBHAM!WCGFMOLE!BFXHO CDB!AH!SOWWFGFDH!SFNFMCQLFDHAM!HOLF!QCODHB! b."KZ_%."aZ_cZ!!.AXT!LIBXMF!OB!MAEFMFS!OD! HTF!."_Z_!XCDHGCM!BFXHOCDZ!!,IBXMF!ADS! ECDFB!AGF!EGAXTOCGASOAMB!b3*cR!F]HFDBCG!XAG QO!GASOAMOB!EGFNOB!b.&*cR!F]HFDBCG! SOPOHCGIL!XCLLIBOB!b.'&cR!F]HFDBCG!SOPOHCGI L!MAHFGAMOB!b.'-cR!WMF]CG!XAGQO!GASOAMOB! b:&*cR!WMF]CG!XAGQO!IMDAGOB!b:&7cR!WMF]CG !SOPOHCGIL!QGCWIDSIB!XAQIH!WASOAMF!b:'0cR! QCMMOXOB!MCDPIB!b0-cR!ADS!QG CDAHCG!HFGFB!b01c!LIBXMFB!ADS!IMDA!b7c!ADS!GASOIB!b*c! ECDFBZ!!*FQGODHFS!WGCL! REE%7#5)='I"=%'S"&'+56%&'J*&5 7D'1@%=%#)='!*E6=%'+%8%="?,%7# g! $6*CIGJF!5*R!)FCGPFB!2+R!2FA4!9*R!1A QBXCHH!25R!,X,ADIB!,1R!)CMSTALFG!'5R!! 2[ADBCD!,2!ADS!9AGWF!3'g!&CQ4GOPTH!=>>_R![OHT!QFGLOBBOCD!WGCL!.MBFNOFGZ!

PAGE 34

! KU! ! ! :OPIGF!"%UZ! +56%&' LIHADHB!BTC[!BJFMFHAM!LIBXMF!T4QCQMABOAZ +c!'FBLOD!OLLIDCTOBHCXTFLOBHG4! CW!XCDHGCM!ADS!LIHADH!HGADBNFGBF!SOBHAM!WC GFMOLE!BFXHOCDB!AH!SOWWFGFDH!SFNFMCQLFDHAM! HOLF!QCODHB!b."KZ_%."aZ_cZ!!2XAMF!EAGB!F YIAM!=_>!qLZ!!3%&c!Z>_Z!!,IBXMF! AEEGFNOAHOCDB!AGF!MOBHFS!OD!:OPZ!=!MFPFDSZ!!*FQGODHFS!WGCL! REE%7#5)='I"=%'S"&'+56%&' J*&57D'1@%=%#)='!*E6=%'+%8%="?,%7# g!$6*CIGJF!5*R!)FCGPFB!2+R!2FA4!9*R!1AQBXCHH! 25R!,X,ADIB!,1R!)CMSTALFG!'5R!!2[AD BCD!,2!ADS!9AGWF!3'g!&CQ4GOPTH!=>>_R! [OHT!QFGLOBBOCD!WGCL!.MBFNOFGZ!!! !

PAGE 35

! K_! ! ! ! :OPIGF!"%_Z!+EDCGLAM!L4CWOEFG! LCGQTCMCP4!ADS!CGPADO\AHOCD!OD! +56%&' LIHADHBZ!!+%3c!&CDHGCM! ADS!LIHADH!MOHHFGLAHF!F]HFDBCG!XAGQO!GASOAMOB !HGADBNFGBF!BFXHOCDB!b+c!CG!HCDPIF!BAPOHHAM! BFXHOCDB!3c!BHAODFS![OHT!ADHOECS4!BQFXOW OX!WCG!SFBLOD!AH!."_Z_!ADS!."`Z_Z!!'+0(! bEMIFc!BHAODB!DIXMFOZ!!(DBFHB!BTC[!GF QGFBFDHAHONF!TOPTFG!LAPDOWOXAHOCD!OLAPFB!CW! HCDPIFBZ!!+MM!BXAMF!EAGB!F YIAM!=_!qLZ!!*FQGODHFS!WGCL! REE%7#5)='I"=%'S"&'+56%&'J*&57D' 1@%=%#)='!*E6=%'+%8%="?,%7# g!$6*CIGJF!5*R!)FCGPFB!2+R!2FA4!9*R!1AQBXCHH!25R! ,X,ADIB!,1R!)CMSTALFG!'5R!!2[ADBCD! ,2!ADS!9AGWF!3'g!&CQ4GOPTH!=>>_R![OHT! QFGLOBBOCD!WGCL!.MBFNOFGZ! !

PAGE 36

! Kd! ! :OPIGF!"%dZ!(DXGFABFS!AQCQHCBOB!OD! +56%&' LIHADH!BJFMFHAM!LIBXMFZ!!+c!Z>_Z! 3%&c!&MFANFS!XABQABF%K!bGFSc!ADS!L4CBOD! TFAN4!XTAOD!bPGFFDc!XCOLLIDCTOBHCXTFLOBH G4!CD!."_Z_!XCDHGCM!b3c!ADS!LIHADH! MOHHFGLAHF!b&c!HGAD BNFGBF!SOBHAM!WCGFMOLE! BFXHOCDB!b">ecZ!!(DBFH!bU>ec!BTC[B!TOPTFG! LAPDOWOXAHOCD!CW!A!LIHADH!BFXHOCDZ!!'c!&M FANFS!XABQABF%K!bGFSc!ADS!L4CBOD!TFAN4! XTAOD!bPGFFDc!XCOLLIDCTOBHCXTFLOBHG4!CD! ."dZ_!LIHADH!BAPOHHAM!SOBHAM!WCGFMOLE! BFXHOCDZ!!#IXMFO!AGF!BHAODFS![OHT !'+0(!bEMIFcZ!!*FQGODHFS!WGCL! REE%7#5)='I"=%'S"&' +56%&'J*&57D'1@%=%#)='!*E6=%'+%8%="?,%7# g!$6*CIGJF!5*R!)FCGPFB!2+R!2FA4!9*R! 1AQBXCHH!25R!,X,ADIB!,1R!)CMSTALFG!'5R! !2[ADBCD!,2!ADS!9AGWF!3'g!&CQ4GOPTH! =>>_R![OHT!QFGLOBBOCD!WGCL!.MBFNOFGZ!!!

PAGE 37

! K`! ! :OPIGF!"%`Z!'FXGFABFS!LIBXMF!LO* #+B!ADS!ODXGFABFS!AQCQHCBOB!CW! +56%& !LIHADH!LIBXMF!XFMMB! 57' 85#&" Z!!+c *#+!EMCH!ADAM4BOB!CW! +56%&' XCDSHOCDAM!,'.*!b%!&GFc!ADS!&GF%ODWFXHFS! +56%&' XCDSOHOCDAM!,'.*!bl!&GFc!WOEGCEMABH!b%! %FBHGASOCMc!ADS!LIBXMF!XFMM!bl! % FBHGASOCMR!=U!TGBZc!XIMHIGFBZ !!LO*%=>d!b[TOXT!OB!ODSIXFS !E4!,4CS%.*!F]QGFBBOCDc!ADS! LO*%"d!b[TOXT!OB!XCDBHOHIHONFM4!F]QGFBBFS!OD! HTFBF!XFMMB!ADS!XAD!EF!IBFS!AB!A!MCASODP! XCDHGCM!WCG!WOGBH!H[C!MADFBc![FGF!ADAM4\FS !E4!BHGOQQODP!ADS!BFYIFDHOAM!T4EGOSO\AHOCD!CW! HTF!BALF!EMCHZ!3c!0TABF%XCDHGABH!OLAPFB!CW!IDWO]FS! +56%&' XCDSOHOCDAM!,'.*!b%!&GFc! ADS!&GF%ODWFXHFS! +56%&' XCDSOHOCDAM!,'.*!bl!&GFc!XFMM BR!WCMMC[ODP!?d!TCIGB!OD!',! [OHT!CG![OHTCIH! %FBHGASOCMR!AB!ODSOXAHFSZ!">e!LAPDO WOXAHOCDZ!!&c!:MC[!X4HCLFHGOX! ADAM4BOB!CW! +56%&' XCDSOHOCDAM!,'.*!b%!&GFc!ADS!&GF%ODWFXHFS! +56%&' XCDSOHOCDAM! ,'.*!bl!&GFc!XFMMBZ '' &FMMB![FGF!XIMHIGFS!OD!', ![OHT!blc!CG![OHTCIH! %FBHGASOCM!b%c! WCG!=U!TCIGBR!AB!ODSOXAHFSZ!!&FMMB![FGF!HG4QB ODO\FSR!BHAODFS![OHT! +DDF]OD!/%:(1&!ADS! QGCQOSOIL!ADS!BCGHFS!ADS!YIADHOWOFS!E4!:+ &2Z!!2TC[D!AGF!BILLAG4!SAHA!CW!HTF! QFGXFDHAPF!CW!QCBOHONFM4%BHAODFS!XFMMBR!GFQGFBFDHODP!HTGFF!ODSFQFDSFDH!HGOAMBZ!rf! 0o>Z>_R!rrf!0o>Z>"!ADS!rrrf!0o>Z>>"Z!!*FQGODHFS!WGCL! REE%7#5)='I"=%'S"&'+56%&' J*&57D'1@%=%#)='!*E6=%'+%8%="?,%7# g!$6*CIGJF!5*R!)FCGPFB!2+R!2FA4!9*R!1AQBXCHH! 25R!,X,ADIB!,1R!)CMSTALFG!'5R!!2[AD BCD!,2!ADS!9AGWF!3'g!&CQ4GOPTH!=>>_R! [OHT!QFGLOBBOCD!WGCL!.MBFNOFGZ!!

PAGE 38

! Ka! :OPIGF!"%aZ!'FXGFABFS!XF MM!QGCMOWFGAHOCD!OD! +56%&' LIHADH!BJFMFHAM!LIBXMF Z!!+c!0TCBQTCTOBHCDF!9K! b09Kc!OLLIDCTOBHCXTFLOBHG4!CW!."KZ_!ADS! "UZ_!XCDHGCM!ADS!LIHADH!MOHHFGLAHF!SOBHAM! WCGFMOLE!XGCBB!BFXHOCDBZ!!+GGC[TFASB!QCODH! HC!09K!QCBOHONF!DIXMFO![OHTOD!BJFMFHAM! LIBXMFBZ!!(DBFH!BTC[B!A!TOPTFG!LAPDOWOXAHOCD!CW!AD!AGGC[TFAS!ADS!A!09K!QCBOHONF! DIXMFIBZ!!2XAMF!EAGB!FYIAM!">>!qLZ!!3c! Z>_Z!!*FQGODHFS!WGCL! REE%7#5)='I"=%'S"&'+56%&' J*&57D'1@%=%#)='!*E6=%'+%8%="?,%7# g!$6*CIGJF!5*R!)FCGPFB!2+R!2FA4!9*R!1AQBXCHH! 25R!,X,ADIB!,1R!)CMSTALFG!'5R!!2[AD BCD!,2!ADS!9AGWF!3'g!&CQ4GOPTH!=>>_R! [OHT!QFGLOBBOCD!WGCL!.MBFNOFGZ!

PAGE 39

! K?! ! ! ! :OPIGF!"%?Z! +56%&' LIHADHB!BTC[!AEFGGADH!F]QGFBBOCD!CW!L 4CPFDOX!WAXHCGBZ!!(LLIDCEMCH!ADAM4BOB! CW!."aZ_!XCDHGCM!bDsKc!ADS!LI HADH!bDsKc!HCDPIF!F]HGAXHB! [OHT!ADHOECSOFB!BQFXOWOX!WCG! ,4C'R!,4CPFDODR!L4CBOD!TFAN4!XTAOD!WABH!OBCWCGL!ADS!SFBLODZ!!)AQST![AB!HTF! MCASODP!XCDHGCMZ!!*FQGODHFS!WGCL! REE%7#5)='I"=%'S"&'+56%&'J*&57D'1@%=%#)='!*E6=%' +%8%="?,%7# g!$6*CIGJF!5*R!)FCGPFB!2+R!2FA4 !9*R!1AQBXCHH!25R!,X,ADIB!,1R! )CMSTALFG!'5R!!2[ADBCD!,2!ADS!9AGWF!3' g!&CQ4GOPTH!=>>_R![OHT!QFGLOBBOCD!WGCL! .MBFNOFGZ! !

PAGE 40

! U>! &9+01.*!=! &$,3(#+1$*(+-!-$22!$:!,3#-"!+#'!', 08%-(#8.'!).#.!.e0*.22($#!:+(-2! 1$!,$'.-!&$#).#(1+-!,;$1$#(&!';21*$09;! Introduction Adult-Onset Myotonic Dystrophy 1TF!L4CHCDOX!S4BHGCQTOFB!AGF!A!PGCIQ!CW!DF IGCLIBXIMAG!SOBFABFB!HTAH!AGF!XAIBFS!E4! LOXGCBAHFMMOHF!F]QADBOCDB!OD!HTF!DCD%XCSODP!GFPO CDB!CW!H[C!SOWWFGFDH!PFDFBR!ADS!ECHT!SOBFABFB! BTC[!AD!AIHCBCLAM!SCLODADH!QAHHFGD!CW!ODTFGO HADXFZ!!,4CHCDOX!'4BHGCQT4!H4QF!"!b',"c!OB! FBHOLAHFS!HC!AWWFXH!"f`>>>!ODSONOSIAMB!LAJODP! OH!HTF!LCBH!XCLLCD!WCGL!CW!ASIMH!LIBXIMAG! S4BHGCQT4!ADS!HTF!BFXCDS!LCBH!QGFNAMFDH!CW!AM M!HTF!LIBXIMAG!S4BHGCQTOFBR!EFTODS!'IXTFDDF! ,IBXIMAG!'4BHGCQT4!b','c!b?>cZ!!VTOMF!,4CHCDOX! '4BHGCQT4!H4QF!=!b',=c!OB!MFBB!QGFNAMFDHR! HTF!TAMMLAGJ!WFAHIGFB!CW!',=!AGF!NFG4!BOLOMAG! HC!',"!b?"cZ!!1TF!XMODOX AM!QGFBFDHAHOCD!XCLLCD! HC!',"!ADS!',=!ODXMISFB!XAHAGAXHBR!BJFMFHAM!LI BXMF![FAJDFBBh[ABHODPR!A DS!L4CHCDOA!bSFMA4!OD! LIBXMF!GFMA]AHOCD!AWHFG!XCDHGA XHOCDcR!ADS!XAGSOAX!SFWFXHBZ! !',"!OB!XAIBFS!E4!A!b&1)c D !GFQFAH!F]QADBOCD!OD!HTF!K6%I DHGADBMAHFS!GFPOCD!bKt%71*c!CW! HTF! +!/0 !PFDF!b:OPZ!=%"+c!b?=%?_cZ!!9FAMHT4!ODSONO SIAMB!H4QOXAMM4!TANF! EFH[FFD!_!ADS!K_!&1)! GFQFAHB!b&1)c _%K_ ![TOMF!',"!QAHOFDHB!TANF!AD!AMMFMF![OHT!b&1)c K_%K>>> Z!!!1TFBF!GFQFAHB!AGF! IDBHAEMF!ADS!AGF!XAQAEMF!CW!F]QADSODP!OD!BIXXFBBONF! PFDFGAHOCDBR!ADS!HTF!BFNFGOH4!CW!HTOB!SOBFABF! XCGGFMAHFB!SOGFXHM4![OHT!HTF!BO\F!CW !HTF!F]QADBOCD!ADS!ODSOGFXHM4![ OHT!HTF!APF!CW!CDBFHZ!!1TF!LCBH! BFNFGFM4!AWWFXHFS!',"!QAHOFDHB!TANF!b&1)c u"R>>> !ADS!QGFBFDH![OHT!A!L4GOAS! CW!XCDPFDOHAM!SFWFXHBZ!!!! !',=!GFBIMHB!WGCL!A!BOLOMAG!LIHAHOCDR!A!b&&1)c D !F]QADBOCD!OD!ODHGCD!"!CW!HTF! 9(:; !PFDF! b:OPZ!=%"3c!b?dcZ!!14QOXAMM4R!b&&1)c ""%`_! AMMFMFB!AGF!QGFBFDH!OD!HT F!DCGLAM!QCQIMAHOCD![TOMF!',=! ODSONOSIAMB!XAGG4!b&&1)c `_%""R>>>! AMMFMFBZ!!b&&1)c D !AGF!IDBHAEMF!ADS!F]QADS!EFH[FFD!PFDFGAHOCDB! EIH!DC!XCGGFMAHOCD!EFH[FFD!ODXGFABFS!GFQFAH!DILEF G!ADS!SOBFABF!BFNFGOH4!TAB!EFFD!CEBFGNFS!DCG! TANF!HTFGF!EFFD!AD4!GFQCGHFS!XABFB!CW!XCDPFDOHAM!',=Z!

PAGE 41

! U"! RNA Dominance Model for Myotonic Dystrophy 1TF!SOBXCNFG4!CW!',"!ADS!',=!LIHAHOCDB!MF S!HC!HTF!WCMMC[ODP!YIFBHOCDf!9C[!SC!BOLOMAG! LIHAHOCDB!OD!DCDXCSODP!GFPOCDB!CW!H[C!XCLQMFHFM4! IDGFMAHFS!PFDFB!GFBIMH!OD!SCLODADHM4!ODTFGOHFS! SOBCGSFGB!HTAH!BTC[!BOLOMAG!XMODOXAM!B4LQHCLB v!!1TF!*#+!'CLODADXF!,CSFM!QGCQCBFB!HTAH!',"! ADS!',=!b&7)c D !ADS!b&&7)c D !F]QADBOCD!L*#+B!QGCNOSF!DCNFM! BQFXOWOX!EODSODP!BOHFB![TOXT! BFYIFBHFG!HTF!,72&-.3-(#'!b,3# -c!QGCHFODB!A[A4!WGCL!HTFOG!DCGLAM!HAGPFHB!GFBIMHODP!OD! ,3#-!MCBB%CW%WIDXHOCD!ADS!SOBFAB F!b:OPZ!=%=cZ!!.NOSFDXF!BIQQCG HODP!HTF!*#+!'CLODADXF!,CSFM! XALF!WGCL!EOCXTFLOXAM!BHISOFB!SFLCDBHGAHODP!HT AH!,3#-!EODS!HC!ADS!XCMCXAMO\F![OHT!',"!ADS! ',=!QAHTCPFDOX!F]QADSFS!GFQFAH!*#+B!b?`%">=cZ!!(LQCGHADHM4R! !<7=!JDCXJCIH!LOXF! b !<7=" .Kh .K c!GFXAQOHIMAHF!BFNFGAM!',"!ADS!', =!TAMMLAGJ!WFAHIGFB!b">KcZ!! !<7=-' LIHADH!LOXF! SFNFMCQ!L4CHCDOAR!XAHAGAXHBR!A DS!XAGSOAX!XCDSIXHOCD!SFWFXHBZ!!:IGH TFGR!,3#-"![AB!OSFDHOWOFS!AB! AD!AMHFGDAHONF!BQMOXODP!WAXHCG !ADS!,EDM"!JDCXJCIH!LOXF!ADS!',!QAHOFDHB!TANF!BOLOMAG!BQMOXODP! AEDCGLAMOHOFB!b">K%">dcZ!!1TFBF!GFBIMHB!SFLCDBHGAH FS!HTAH!,3#-!MCBB!OB!A!QGOLAG4!QAHTCPFDOX! FNFDH!XAIBODP!ASIMH%CDBFH!',!XMOD OXAM!ADS!LCMFXIMAG!SFWFXHBZ!!! Congenital Myotonic Dystrophy (CDM) &',!OB!XAIBFS!E4!HTF!BALF!b&1)cD!F]QADBOCD!WCIDS!OD!ASIMH%CDBFH!',"!b?>cZ!!9C[FNFGR! &',!LADOWFBHB![TFD!HTF!GFQFAH!F]QADBOCD!F]XFF SB!A!HTGFBTCMS!CW!b& 1)cu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b&&7)c u">R>>> !AMHTCIPT!DC!XCDPFDOHAM!SF WFXH!OB!CEBFGNFS!OD! HTFBF!QAHOFDHB!b">`cZ!!

PAGE 42

! U=! 1TOB!BIPPFBHB!HTAH!HTF!OLQAOGFS!FLEG4CDOX!L4CPFDFB OB!ADS!DFIGCPFDFBOB!BFFD!OD!&',!OB!BQFXOWOX! HC!HTF!',"!MCXIBZ! The Dm1 Locus: DMWD DMPK And SIX5 1TF!b&1)c D !LIHAHOCD!CEBFGNFS!OD!&',!OB!MCXAHFS!OD!HTF!K6!71*!CW!HTF! +!/0 !PFDFZ!! (LLFSOAHFM4!IQBHGFAL!CW! +!/0 !MOFB! +!.+ !ADS!SOGFXHM4!SC[DBHGFAL!OB! 1234 Z!!1TFBF!HTGFF! PFDFB!AGF!XMIBHFGFS!HCPFHTFG!OD !k=>JE!CW!PFDCLOX!'#+!ADS![OMM!EF! GFWFGGFS!HC!AB!HTF!',"!MCXIB! b:OPZ!=%KcZ!!1TF!XFMMIMAG!WIDXHOCDB!CW!HTFBF! QGCHFODB!AGF!IDJDC[DR!EIH!FAXT!XCDHAODB!BQFXOWOX! LCHOWB!HTAH!AGF!QGFBFDH!OD !A!NAGOFH4!CW!QGCHFODB![OHT!JDC[D!WIDXHOCDZ!! 1TF!PFDF!IQBHGFAL!CW! +!/0 !OB!DALFS! +!.+ !EFXAIBF!OH!OB!ABBCXOAHFS![OHT!HTF!',"! MCXIB!ADS!OHB!QGCHFOD!QGCSIXH!XCDHAODB !WCIG!HG4QHCQTAD%ABQAGHOX!AXOS!bV' c!GFQFAHBZ!!,CIBF! +,TJ OB!F]QGFBBFS!OD!AMM!ASIMH!HOBBIFBR!EIH !OB!LCBH!QGCLODFDH!OD !EGAOD!ADS!HFBHFB!b">ac Z!!(H!OB!QGFBFDH!OD! AMM!AGFAB!CW!HTF!EGAODR!EIH!OB!FDGOXTFS!OD!HTF! XFMM!ECS4R!DIXMFIBR!ADS!SFDSGOHFB!CW!DFIGCDBZ!!(D!HTF! HFBHFBR! +,TJ !OB!F]QGFBBFS!OD!BQFGLAHCX4HFB!ADS!2FGHC MO!XFMMB!b">?cZ!!!!+ MHTCIPT!HTF!WIDXHOCD!CW! ',V'!GFLAODB!HC!EF!FMIXOSAHFSR!CHTFG!QGCHFODB!TA NF!EFFD!OSFDHOWOFS!HTAH! TANF!WCIG!HC!FOPTH!V'! GFQFAHB!ADS!HTFBF!GFQFAHB!WCMS!ODHC!A!H[OBHFS!QGCQ FMMFG!BHGIXHIGF![TOXT!XGFAHFB!A!BHAEMF!QMAHWCGL! HTAH!XAD!WCGL!XCLQMF]FB![OHT!CH TFG!QGCHFODB!b"">cZ!!V'!GFQFAH !XCDHAODODP!QGCHFODB!TANF!EFFD! BTC[D!HC!WIDXHOCD!OD!BOPDAM!HGADBSIXHOCDR!*#+!QG CXFBBODPR!HGADBXGOQHOCDR!X4HCBJFMFHCD!ABBFLEM4R! XFMM!SONOBOCDR!ADS!NFBOXIMAG!HGAWWOXJODPZ!!! !(LLFSOAHFM4!SC[DBHGFAL!CW! +!.+ !OB! +!/0 Z!!,CIBF! +,?@ !OB!F]QGFBBFS!OD!LCBH!XFMM! H4QFB!EIH!HTF!TOPTFBH!F]QG FBBOCD!CXXIGB!OD!BJFMFHAM!LIBXMFR!TF AGHR!ADS!HTF!BLCCHT!LIBXMF!MODODPB! CW!CGPADB!OD!ASIMHB!ADS!OD!HTF!SFNFMC QODP!MOLEB!ADS!EGAOD!CW!HTF!FLEG4C!b"""R""=cZ! +!/0 FDXCSFB!A!BFGODFhHTGFCDODF!QGCHFOD!JODABF!GFMAHFS! HC!QGCHFOD!JODABFB!&!ADS!+R!ADS!HTF!*TC!WALOM4! CW!JODABFBZ!!(H!TAB!EFFD!BTC[D!HC!QTCBQTCG4MAHF!HTF! !BIEIDOH!CW!HTF!SOT4SGCQ4GOSODF!GFXFQHCG! b'90*cR!L4CBOD!QTCBQTAHABF!b,;01"c!ADS!QTCBQTCMALEAD!b0-#c! 57'85#&"' b""K%""_cZ!!',08!

PAGE 43

! UK! L*#+!OB!AMHFGDAHONFM4!BQMOXFS!ADS!SOWWFGFDH! &%HFGLODO!SOXHAHF!HTF!XFMMIMAG!MCXAMO\AHOCD!ADS! BIEBHGAHF!BQFXOWOXOH4!CW!HTOB!JODABF!b""dcZ!!(B CWCGLB![OHT!A!T4SGCQTCEOX!&% HFGLODIB!AGF!AHHAXTFS! HC!HTF!.*R!HTCBF![OHT!A!T4SGCQTOMOX!&%HFGLODIB!AG F!MCXAHFS!AH!LOHCXTCDSGOA R!ADS!OBCWCGLB![OHT!A! BTCGH!&%HFGLODIB!AGF!X4HCQMALOXZ!!',08!MCXA MO\FB!HC!HTF!DFIGCLIBXIMAG!iIDXHOCD!b#,5c!OD! LIBXMF!ADS!HC!PAQ!iIDXHOCDB!OD!HTF!TFAGH!b""`cZ !!1TFBF!AGFAB!AGF!QCQIMAHFS!E4!DILFGCIB!H4QFB!CW! OCD!XTADDFMB!ADS!B IEBHGAHFB!OSFDHOWOFS! 57'85#&" !AGF!ODNCMNFS!OD!GFPIMAHODP! XAMXOIL!OCD!MFNFMBZ!!1TOB! BIPPFBHB!HTAH!CDF!WIDXHOCD!CW !',08!OB!HC!GFPIMAHF!&A =l !OCD!MFNFMB!OD!NAGOCIB!HOBBIFBZ! 1234 !FDXCSFB!A!TCLFCSCLAOD!HGADBXGOQHOCD!WAXHCG! ADS!OB!MCXAHFS!SOGFXHM4!K6!HC!HTF!b&1)c D OD!HTF! +!/0' K6!71*Z!!+!&Q)!OBMADS!BIGGCIDSB!HTF!b&1)c D !ADS!HTF! 1234 !QGCLCHFG!FMFLFDHB!AGF! MCXAHFS!OLLFSOAHFM4!SC[DBHGFAL! CW!HTOB!GFQFAH!b""acZ!!-OJF! +!.+ !ADS! +!/0 R! 1234 !OB! F]QGFBBFS!OD!BFNFGAM!HOBBIFBg! TC[FNFGR!2(e_!QGCHFOD!SCFB!DCH!A QQFAG!HC!EF!FDGOXTFS!OD!AD4! QAGHOXIMAG!CGPAD!B4BHFL!b""?cZ!! 1234' OB!A!LFLEFG!CW!A!PFDF!WALOM4![TOXT!OB!XTAGAXHFGO\FS!E4!HTF! QGFBFDXF!CW!2O]!ADS!TCLFC!SCLAODBR!ECHT!CW! [TOXT!QMA4!GCMFB!OD! '#+!EODSODP!ADS!QGCHFOD% QGCHFOD!ODHFGAXHOCDB!b"=>cZ!!1TF!2(e_!TCLF CSCLAOD!AXHB!AB!A!BF YIFDXF%BQFXOWOX!'#+!EODSODP! SCLAOD!ADS!HTF!2O]!SCLAOD!OB!GFYIOGFS!WCG!HTF!WC GLAHOCD!CW!A!BHAEMF!TCLCSOLFGOX!XCLQMF]!HTAH! FDTADXFB!'#+!EODSODPZ!!3CHT!SCLAODB!AGF!GF YIOGFS!WCG!2(e_!EODSODP!HC!HTF!+*.R!A! HGADBXGOQHOCDAM!GFPIMAHCG4!FMFLFDH!OD!HTF!#A%8!+10ABF!"!PFDF!b PQ/-Pc!b"="cZ!!2(e_!AMBC! BTC[B!EODSODP!BQFXOWOXOH4!WCG!L4CPFDOX!FDTADXODP!W AXHCG!K!LCHOWB!b,.:Kc![TOXT!AGF!QGFBFDH!OD!A! NAGOFH4!CW!BJFMFHAM!LIBXMF!BQFXOWOX!GFPIMAH CG4!PFDFB!ODXMISODP!L4CPFDOD!b"==cZ!1C!IDXCNFG! QCHFDHOAM!HAGPFHB!CW!2(e_R!F]QG FBBOCD!QGCWOMODP![AB!XAGGOFS!CIH!CD! 0"?!XFMMB!HTAH!CNFGF]QGFBBFS! HTOB!WAXHCG!b"=KcZ!!1[FDH4!CDF!PFDFB!BTC[FS!OD XGFABFS!F]QGFBBOCD!OD!HTF!XFMMB!HTAH!CNFGF]QGFBB! 2(e_R!HTF!LAiCGOH4!CW![TOXT!AGF!F]QGFBBFS!OD!BJFMFH AM!LIBXMF!ADS!EGAODZ!!1TOB!OLQMOFB!HTAH!CDF!CW! HTF!WIDXHOCDB!CW!2(e_!OB!HC!GF PIMAHF!LIBXMF!ADS!EGAOD!WIDXHOCDZ!

PAGE 44

! UU! !+MHTCIPT!HTF!QGFXOBF!XF MMIMAG!WIDXHOCDB!CW! +!.+ R! +!/0 !ADS! 1234 !TANF!4FH!HC!EF! GFNFAMFSR!FAXT!OB!F]QGFBBFS!OD!HTF!HOBBIFB!HT AH!AGF!AWWFXHFS!OD!&' ,!QAHOFDHB!ADS!AEFGGADH! F]QGFBBOCD!CW!HTFBF!QGCHFODB!LA4!XCDHGOEIHF! HC!HTF!LADOWFBHAHOCD!CW!HTOB!SOBFABFZ! The Dmpk Expanded CTG Repeat alters Dm1 Locus Chromatin Conformation (H![AB!CGOPODAMM4!T4QCHTFBO\FS! HTAH!HTF!XMODOXAM!QG FBFDHAHOCD!CEBFGNFS!OD!',"!GFBIMHB!WGCL! SFXGFABFS!F]QGFBBOCD!CW!HTF!',"!MCXIBZ!!2IQQCGH! WCG!HTOB!LCSFM!XALF!WGCL!BHISOFB!BTC[ODP!HTAH! A!b&1)c D !F]QADBOCD!AMHFGB!HTF!XTGCLA HOD!BHGIXHIGF!BIGGCIDSODP!HTF! GFQFAHZ!!.MFXHGCD!LOXGCBXCQ4! ADS! 57'85#&" !DIXMFCBCLF!GFXCDBHOHIHOCD!A BBA4B!GFNFAMFS!HTAH!HTFGF![AB!A!SOGFXH!XCGGFMAHOCD! EFH[FFD!HTF!DILEFG!CW!b&1)c D !GFQFAHB!ADS!HTF!FWWOXOFDX4!CW!DIXMFCBCLF!WCGLAHOCD!b"=UcZ!!(D! ASSOHOCDR!MCDP!&1)!GFQFAHB!XAD!BFGNF!AB!DIXMFCBCLF!QCBOHOCDODP!FMFLFDHB!HTAH!AMHFG!MCXAM! XTGCLAHOD!XCDWCGLAHOCDR!LAJODP!OH!MFBB!AXXFBBOEMF! HC!DIXMFABF!SOPFBHOCDR! A!XTAGAXHFGOBHOX!CW! TFHFGCXTGCLAHOD!b"=_cZ!!:IGHTFGLCGFR!DCGLAM!b&1)c D !AGF!WMADJFS!E4!&1&:!EODSODP!BOHFB!HTAH!AGF! LFHT4MAHFS!CD!&',!LIHADH!AMMF MFB!ADS!QGFNFDH!&1&:!EODSODP!b "=dcZ!!&1&:!OB!AD!ODBIMAHCG! EODSODP!QGCHFOD!HTAH!WIDXHOCDB!HC!QGFNFDH!TFHFGC XTGCLAHOD!BQGFAS!CG!HC !EMCXJ!FDTADXFGB!WGCL! AXHODP!CD!DFAGE4!QGCLCHFGB!b"=`cZ!!1TFGFW CGFR!AMMFMFB!XCDHAODODP!F]QADSFS!b&1)c D !LIHAHOCDB! LA4!EF!OD!LCGF!CW!TFHFGCXTGCLAHOD!XCDWCGLA HOCD!HTAH!GFBIMHB!OD!SFXGFABFS!F]QGFBBFS!WGCL! +!% MODJFS!PFDFBZ!! !(D!CGSFG!HC!HFBH!HTF!',"!MCXIB!MCBB%CW%W IDXHOCD!T4QCHTFBOBR!LAD4!MAEB!XCDSIXHFS!A!EAHHFG4! CW!F]QFGOLFDHB!HC!F]ALODF!HTF!FWWFXHB!CW!HTF!b&1)c D !F]QADBOCD!CD!HTF!F]QGFBBOCD!CW! +!.+ R! +!/0 !ADS! 123 Z!!2FNFGAM!GFQCGHB!TANF!SFLCDBHGAHFS! HTAH!',08!ADS!2(e_!*#+!MFNFMB!AGF! GFSIXFS!OD!',"!ADS!&',!HOBBIFB!ADS!XIMHIGFS!XFMMB !AMHTCIPT!CHTFG!BHISOFB!GFQCGH!DC!XTADPF!CG! AD!ODXGFABF!OD!F]QGFBBOCD!b"=a%"K?cZ!1TF!GFBIMHB! CW!HTFBF!F]QFGOLFDHB!GAOBF!HTF!QCBBOEMOH4!HTAH! &',!LA4!GFBIMH!WGCL!TAQMCODBIWWOXOFDX4!CW !CDF!CG!LCGF!CW!HTF!',"!MCXIB!PFDFBZ!

PAGE 45

! U_! Disruption of The Mouse Dm1 Locus (W!HTF!T4QCHTFBOB!OB!HGIF! HTAH!HTF!F]QADSFS!b&1)c D !XAIBFB!GFSIXFS!F]QGFBBOCD!WGCL!HTF! ',"!MCXIB!HTFGFE4!GFBIMHODP!OD!&',R!HTFD!SFXG FABFS!F]QGFBBOCD!WGCL!HTFBF!PFDFB!CIHBOSF!HTF! XCDHF]H!CW!HTF!b&1)c D !BTCIMS!GFXAQOHIMAHF!HTF!QTFDCH4QFB!ABBCXO AHFS![OHT!HTOB!SOBFABFZ!!1C!HFBH! HTOB!T4QCHTFBOBR!PFDF!HAGPFHODP!F]QFGOLFDHB!APAODBH!LCIBF! +,?@' ADS! 15A4 ![FGF!XCDSIXHFSZ!!,OXF! AGF!A!NAMIAEMF!PFDFHOX!HCCM!BODXF!HTF!',"!MCXI B!OB!GFMAHONFM4!XCDBFGNFS!EFH[FFD!TILADB!ADS! LOXF![OHTOD!OD!A!MAGPF!BFPLFDH!CW!B4DHFDOX!'#+!b"U>cZ!!1C!SAHFR!DC! +,TJ !JDCXJCIH!LOXF!TANF! EFFD!PFDFGAHFSZ! +,?@ lh% !ADS! +,?@ %h%! [FGF!PFDFGAHFS!E4!H[C!ODSFQFDSFDH! MAECGAHCGOFB!HC!ODNFBHOPAHF!HTF! FWWFXH!CW!MC[FGFS!MFNFMB!CW!'LQJ!CD!LCIBF! SFNFMCQLFDHZ!!3CHT!PGCIQB!IBFS!TCLCMCPCIB! GFXCLEODAHOCD!OD!LCIBF!FLEG4CDOX!BHFL!XFMMB !HC!GFQMAXF!HTF!WO GBH!BFNFD!F]CDB!CW! +,?@ ![OHT!AD! ADHOEOCHOX!GFBOBHADXF!XABBFHHF!GF BIMHODP!OD!HTF!XCLQMFHF!AEBFDXF!CW!'LQJ!QGCHFOD!OD!JDCXJCIH! TCLC\4PCHFB!b"U"R"U=cZ!!1TFBF!LOXF!AQQFAGFS!HC !EF!QTFDCH4QOXAMM4!DCGLAM!AMHTCIPT!XMCBFG! F]ALODAHOCD!GFNFAMFS!A!LOMS!BJFMFHAM!LIBX MF!L4CQAHT4!XTAGAXHFGO\FS!E4!SFXGFABFS!WCGXF! PFDFGAHOCDR!ODXGFABFS!WOEFG!SFPFDFGAHOCD!ADS!GFPF DFGAHOCD!ADS!MCBB!CW!BAGXCLFGOX!CGPADO\AHOCD! b"U=cZ!!1TOB!BALF!LCIBF!MODF!F]TOEOHFS!XAGSOAX!XCDSIXHOCD!AEDCGLAMOHOFB![TOXT!ODXMISF!WOGBHR! BFXCDSR!ADS!HTOGS!SFPGFF!AH GOCNFDHGOXIMAG!EMCXJ!OD! +,?@ %h%! LOXF!ADS!WOGBH!SFPGFF!TFAGH!EMCXJ!OD! +,?@ lh%! b"UKcZ!!1TF!BFXCDS!MODF!CW!LOXF!BTC[FS!A!NFG4! LOMS!SFXGFABF!OD!BJFMFHAM!LIBXMF!WOEFG! SOALFHFGZ!!+SSOHOCDAMM4R! 57'85#&" !SOWWFGFDHOAHOCD!CW!L4CHIEFB!WGCL! L4CEMABHB!OBCMAHFS!WGCL!HTFBF! ADOLAMB!BIPPFBHFS!AD!AMHFGFS!GFBQCDBF!HC!XA MXOIL!b"U"R"UUcZ!!:IGHTFGLCGFR!AEDCGLAM!BCSOIL! PAHODP![AB!GFQCGHFS!OD!HTF!WOGBH!LCIBF!MODF!ADS!AE DCGLAM!XAMXOIL!X4XMODP!OD!HTF!BFXCDSR!ECHT!OD! OBCMAHFS!XAGSOCL4CX4HFB!b"U_R"UdcZ!!1TF!BIEHMF! SOWWFGFDXFB!CEBFGNFS!OD!HTF!H[C!SOWWFGFDH! +,?@ h%! MODFB!LA4!EF!AHHGOEIHFS!HC!BHGAOD!EAXJPGCIDS!SOWWFGF DXFBZ!!9C[FNFGR!OH!AQQFAGB!FNOSFDH!ECHT!WGCL! HTF! 57'85#&" !ADS!PFDF!HAGPFHODP!BHISOFB!HTAH! +,?@ !OB!ODNCMNFS!OD!GFPIMAHODP! OCD!TCLFCBHABOBZ!!(H!OB!

PAGE 46

! Ud! YIOHF!QCBBOEMF!HTAH!MCBB!CW!',08!XAD!F]QMAOD! BCLF!CW!HTF!B4LQHCLB!ABBCXOAHFS![OHT!',"R! QAGHOXIMAGM4![OHT!GFPAGS!HC! XAGSOAX!XCDSIXHOCD!SFWFXHB!ADS!AEDCGLAM!BJFMFHAM!LIBXMF!QT4BOCMCP4Z!! 9C[FNFGR!OH!OB!AMBC!FNOSFDH!HTAH!HTF!MCBB!CW!'L QJ!AMCDF!SCFB!DCH!GFBIMH!OD !B4LQHCLB!ABBCXOAHFS! [OHT!&',Z! !1C!SFHFGLODF!OW!A!SFXGFABF!OD! 1234 !SCBAPF!XCIMS!F]QMAO D!HTF!',"!ADShCG!&',! QTFDCH4QFR! 15A4 !JDCXJCIH!LOXF![FGF!XGFAHFS!E4!H[C!ODSF QFDSFDH!MAECGAHCGOFB!b"U`R"UacZ!!1TF! WOGBH!PGCIQ!GFQMAXFS!HTF!WOGBH!F]CD!CW! 15A4 ![OHT!HTF! %PAMAXHCBOSABF!PFDF!ADS!HTF!BFXCDS!MODF![AB! XCDBHGIXHFS!E4!GFQMAXODP!AMM!HTGFF! F]CDB![OHT!A!DFCL4XOD!GFBOBHAD XF!XABBFHHFZ!!3CHT!MODFB!CW!LOXF! SFNFMCQFS!CXIMAG!XAHAGAXHB!ADS!B TC[FS!FMFNAHFS!'LQJ!L*#+!MFNFMB !OD!TFAGH!ADS!F4F!HOBBIFB!CW! 15A4 lh% !ADS! 15A4 %h%! LOXFZ!!2(e_![AB!AMBC!BTC[D!HC!EODS!HC! PQ/-P! 57'85#&" !ADS!HTFGFWCGF!L*#+! MFNFMB!CW!+HQ"+"![FGF!F]ALODFS!OD!HTF! 15A4 !JDCXJCIHBZ!!1TF!WOGBH! JDCXJCIH!MODF!BTC[FS!DC! XTADPF!OD!+HQ"A"!L*#+!MFNFMB![TOMF!HTF!BFXCDS! MODF!SOBQMA4FS!+HQ"A"!L*#+!MFNFMB!HTAH![FGF! ODNFGBFM4!QGCQCGHOCDAM!HC! 15A4 !SCBAPFZ!!1TF!GFABCD!WCG!HTOB!SOBXGFQADX4!OB!IDJDC[DZ!!1TF!BFXCDS! MODF!CW! 15A4 %h%! LOXF!BTC[FS!LAMF!BHFGOMOH4!ADS!A!SFXGFABF!OD!HFBHOXIMAG!LABB![OHT!APF![TOXT!LA4! GFBIMH!WGCL!SFXGFABFS! X%8OH!MFNFMB!b"U?cZ!! 15A4 lh% !ADS! 15A4 %h%! LOXF!WGCL!HTOB!MODF![FGF!AMBC!GFQCGHFS! HC!TANF!XAGSOAX!XCDSIXHOCD!SFWFXHB! b"_>cZ!!1TFBF!SAHA!BIPPFBH!HTAH! 15A4 !WIDXHOCDB!OD!HTF! SFNFMCQLFDH!CW!HTF!MFDBZ!(H!OB!OLQCGHADH!HC!DCHF! HTAH!HTF!DIXMFAG!XAHAGAXHB!CEBFGNFS!OD!HTFBF!LOXF! SOWWFG!WGCL!HTF!BIEXAQBIMAG!SIBH%MOJ F!CQAXOHOFB!CEBFGNFS!OD!',"!QAHOFDHBZ!!!1TFBF!GFBIMHB!AMBC! QGCNOSF!FNOSFDXF!HTAH!A!SFXGFABF!OD! 1234 !F]QGFBBOCD!LA4!XCDHGOEIHF! HC!HTF!XAGSOAX!XCDSIXHOCD! ADS!T4QCPCDASOBL!SFWFXHB!CEBFGNFS!OD!ASIMH%CDBFH! ',"!QAHOFDHBZ!!9C[FNFGR!2O]_!MCBB![AB!DCH! GFQCGHFS!HC!XAIBF!AD4!XCDPFDOHAM!AEDCGLAMOHOFBZ! !(D!BILLAG4R!HTFBF!HGADBPFDOX!ADOLAM!BHISO FB!ODSOXAHF!HTAH!SF XGFABFS!F]QGFBBOCD!CW! +!/0 ADS! 1234 !LA4!AXXCIDH!WCG!BCLF!CW!B4LQHCLB!HTAH!AG F!ABBCXOAHFS![OHT!ASIMH%CDBFH!',"Z!;FHR!OH!

PAGE 47

! U`! AQQFAGB!XMFAG!HTAH!A!SFXGFABF!OD!SCBAPF!OD!FAXT !CW!HTFBF!PFDFB!AMCDF!SCFB!DCH!GFBIMH!OD!HTF! DFCDAHAM!SFWFXHB!CEBFGNFS!OD!&',Z!! Results Normal Neonatal Phenotype for Mbnl1;Dmpk and Mbnl1;Six5 Double Knockout Mice !<7=" .Kh .K !LOXF!MAXJ!,EDM"!OBCWCGLB!HTAH!EODS !F]QADSFS!b&c&7)!F]QADBOCD!*#+B!ADS! SFNFMCQ!BFNFGAM!WFAHIGFB!CW!ASIMH%CDBFH!', !ODXMISODP!L4CHCDOA!ADS!BJFMFHAM!LIBXMF! TOBHCQAHTCMCP4R!XAHAGAXHB!ADS!AMHFGDAHONF!BQMOXO DP!SFWFXHB!AGCIDS!BO]![ FFJB!CW!APF!b">KcZ!! 9C[FNFGR!HTFBF!LOXF!AQQFAG!DCGLAM!IDHOM!d%a![FFJB!CW!APF![TFD!L4CHCDOA!OB!WOGBH!CEBFGNFSZ!! 3FXAIBF!QGFNOCIB!BHISOFB!TANF!B IPPFBHFS!HTAH!MCBB!CW!',08!CG! +!/0 %MODJFS!PFDFB!LOPTH!EF! ODNCMNFS!OD!HTF!XCDPFDOHAM!WCGL!CW! HTOB!SOBFABFR![F!IBFS!A!PFDFHOX !AQQGCAXT!HC!HFBH!HTF!T4QCHTFBOB! HTAH!&',!GFBIMHB!WGCL!,3#-!MCBB!XCLEODFS![ OHT!GFSIXFS!',08!ADS!2(e_!QGCHFOD!MFNFMBZ!! !<7=" RKU RK >+,?@ FUF !ADS! !<7=" RKU RK >15A4 FUF !LOXF![FGF!PFDFGAHFS!E4!XGCBBODP! !<7=VU RK TFHFGC\4PCHFB!HC! +,?@ FUF !ADS! 15A4 FUF !TCLC\4PCIB!JDCXJCIHBZ!!VF!QFGWCGLFS!PFDCLOX!'#+! PFDCH4QODP!ADS!*1%0&*!HC!NFGOW4!HTAH!,EDM"R! 'LQJ!ADS!2O]_!MFNFMB![FGF!XCLQGCLOBFS!OD! LIHADH!LCIBF!MODFB!b:OPZ!=%U!ADS!SAHA!DCH!B TC[DcZ!!,EDM"!ADS!'LQJ!L*#+B![FGF!IDSFHFXHAEMF! OD! !<7=" RKU RK >'+,?@ FUF !LOXF!AB![FGF!,EDM"!ADS!2O]_!OD! !<7=" RKU RK >'15A4 FUF ![TFD!XCLQAGFS!HC! XCDHGCM!LIBXMFZ!!!!!!!!!!!!!! &',!XMODOXAM!LADOWFBHAHOCDB! ODXMISF!DFCDAHAM!GFBQ OGAHCG4!ODBIWWOXOFDX 4R!WFFSODP!QGCEMFLB! ADS!WAOMIGF!HC!HTGONFZ!!9C[FNFGR! !<7=" RKU RK >'+,?@ FUF !ADS! !<7=" RKU RK >'15A4 FUF !SCIEMF!JDCXJCIHB! AQQFAGFS!CNFGHM4!DCGLAM!AH!EOGHTR!MONFS!HC!EF!u" !4FAG!CW!APF!ADS![FGF!ODSOBHODPIOBTAEMF!WGCL! !<7=" RKU RK !BODPMF!JDCXJCIHB!OD!HFGLB!CW!QFGODAHAM!NO AEOMOH4R!MCDPFNOH4!ADS!LCEOMOH4!AB!ABBA4FS! E4!XAPF!EFTANOCGZ!!1TFGFWCGFR!MCBB!CW!FOHTFG!'LQJ !ADShCG!2O]_!F]QGFBBOCD !SCFB!DCH!BOPDOWOXADHM4! AMHFG!HTF!,EDM"!JDCXJCIH!QTFDCH4QFZ!!!!!!!!!!

PAGE 48

! Ua! Combinatorial Mbnl1, Dmpk And Six5 Knockouts Fail to Model CDM-Relevant Delays in Muscle Development 2FNFGAM!BHISOFB!TANF!BIPPFBHFS!HTAH !&1)!GFQFAH!F]QADBOCDB!OD!HTF! +!/0 !PFDF!TANF!A! GFQGFBBONF!PMCEAM!FWWFXH!CD!HT F!F]QGFBBOCD!CW!PFDFB!AH!HTF! +!/0 !MCXIB!ADS!HTOB!XCCGSODAHF!MCBB!OB! GFBQCDBOEMF!WCG!HTF!&',!QTFDCH4QFZ!!&',!LADO WFBHB!SIGODP!FLEG4CPFDFBOB!ADS!OB!HTCIPTH!HC! GFBIMH!WGCL!SFMA4B!OD!L4CPFDFBOB!ADS!DFIGCPFDFBOBZ !!1TIBR![F!DF]H!ODNFBHOPAHFS!HTF!QCBBOEOMOH4! HTAH!,3#-"!MCBB!HCPFHTFG![OHT! MCBB!CW!ECHT!',08!ADS!2(e_![CIMS!GFBIMH!OD!&',%ABBCXOAHFS! FLEG4CDOX!ADS!DFCDAHAM!LIBXMF!SFWFXHBZ!!VOMS!H4QFR! !<7=" RKU RK >'+,?@ FUF !ADS! !<7=" RKU RK >' 15A4 FUF !MAHF%BHAPF!FLEG4CB!ADS!DF[ECGD!QIQB![FGF! QTFDCH4QOXAMM4!BOLOMAG!ADS!SOS!DCH!AQQFAG!HC! TANF!PGCBB!ADAHCLOXAM!SFWFXHB!b: OPZ!=%_!ADS!SAHA!DCH!BTC[DcZ!!2J FMFHAM!LIBXMF!SFNFMCQLFDH![AB! F]ALODFS!OD!SFHAOM!EFXAIBF!OH!OB!HTF!LCBH![FMM! XTAGAXHFGO\FS!AEDCGLAMOH4!OD!&',Z!!-AHF%PFBHAHOCD! FLEG4CB!ADS!DFCDAHFB!BTC[!OLLAHIGF!BJFMFHAM! LIBXMF![OHT!AD!IDIBIAM!AEIDSADXF!CW!L4CHIEFBR! HTOD!SOBCGPADO\FS!L4CWOEFGB!A DS!XFDHGAMO\FS!DIXMFO!b"_"%"_KcZ ,4CPFDFBOB!OB!BOLOMAG!OD!TILADB! ADS!LOXF![TFGF!LIBXMF!QGFXIGBCGB!ODOHOAMM4!SOWWF GFDHOAHF!ADS!WIBF!HC!WCGL!FMCDPAHFS!L4CHIEFB! [OHT!DIXMFO!QCBOHOCDFS!OD!HTF!XFDHFG!b"_UcZ!!+H !MAHFG!FLEG4CDOX!BHAPFB!L4CWOEFGB!LAHIGFR! XCDHGAXHOMF!QGCHFODB!AXXILIMAHF!OD!HTF!L4CWOEFG! XFDHFG!ADS!DIXMFO!LOPGAHF!HC!HTF!QFGOQTFG4Z!!VF! F]ALODFS!BJFMFHAM!L4CWOEFG!SFNFMCQLFDH!OD! !<7=" RKU RK >'+,?@ FUF !ADS! !<7=" RKU RK >'15A4 FUF !LOXF! E4!SFBLOD!OLLIDCTOBHCXTFLOBHG4!CD!WCGFMOLE!XGC BB%BFXHOCDB!CW!."_Z_!ADS!."aZ_!FLEG4CB!b:OPZ! =%_cZ!!'FBLOD!OB!A!BJFMFHAM!LIBXMF!BHGIXHIGAM!QGCH FOD!ADS![AB!IBFS!HC!BQFXOWOXAMM4!MAEFM!L4CWOEFGBZ!! .LEG4CDOX!b.c"_Z_![OMS!H4QFR! !<7=" RKU RK >'+,?@ FUF !ADS! !<7=" RKU RK >'15A4 FUF !LIHADH!LIBXMFB! [FGF!ODSOBHODPIOBTAEMF!ADS!XCDBOBHFS!QGOLAGOM4 !CW!OLLAHIGF!L4CWOEFGB!HTAH!QGFBFDHFS!AB! L4CHIEFB![OHT!XFDHGAMO\FS!DIX MFO!b:OPZ!=%_+R!&!ADS!.cZ!!34!."aZ_R!AMM![OMS!H4QFR! !<7=" RKU RK g! +,?@ VUF >'15A4 lh% )7J'!<7=" RKU RK >'+,?@ FUF !ADS! !<7=" RKU RK >'15A4 FUF !L4CHIEFB!SFNFMCQFS!ODHC! LAHIGF!L4CWOEFGB![OHT!BHGCDP!SFBLOD!ADS!L4CBOD !TFAN4!XTAOD!b,9&c!BHAO DODP!OD!HTF!XFDHFG!CW!

PAGE 49

! U?! L4CWOEFGB!ADS!HTF!DIXMFO![FGF!QCBOHOCDFS!DFAG!HT F!BAGXCMFLLA!b:OPZ!=%_+R!'R!:!ADS!SAHA!DCH! BTC[DcZ!!1TFBF!GFBIMHB!SFLCDBHGAHFS!HTAH!MCBB! CW!,EDM"!F]QGFBBOCD!XCLEODFS![OHT!FOHTFG!'LQJ! ADShCG!2O]_!TAQMCODBIWWOXOFDX4! CG!AEMAHOCD!SCFB!DCH!SFMA4!BJFMFH AM!L4CWOEFG!SFNFMCQLFDH!ADS! LAHIGAHOCDZ!!!!!!!!!!!!!!!!! Absence of Limb Deformities in Late Stage Knockout Embryos +DCHTFG!SOBHODPIOBTODP!WFAHIGF! CW!&',!DF[ECGDB!OB!HTF!CX XIGGFDXF!CW!HAMOQFBR!XCLLCDM4! GFWFGGFS!HC!AB!XMIEWCCH!b"__cZ!!1A MOQFB!LA4!GFBIMH!WGCL!XCDPFDOHAM! GFBHGOXHOCD!CW!iCODH!LCNFLFDHR! CG!AGHTGCPG4QCBOBR![TOXT!CXXIGB!SIF!HC!SFNFM CQLFDHAM!XTADPFB!HTAH!OLQAOG!bDFIGCLIBXIMAG! [FAJDFBBc!CG!GFBHGOXH!bAEDCGLAM!WFHAM!QCBOHOCDO DPc!LCNFLFDH!OD!IHFGC!b'GZ!5CTD!'A4R!QFGBCDAM! XCLLIDOXAHOCDcZ!!1C!SFHFGLODF!OW!'LQJ!ADShCG! 2O]_!MCBB!OD!ASSOHOCD!HC! ,EDM"!SFWOXOFDX4![AB! ABBCXOAHFS![OHT!MOLE!SFWCGLOHOFB!OD!&',![F!XCLQ AGFS!BJFMFHAM!QGFQAGAHOCDB!CW!MAHF!PFBHAHOCD! [OMS!H4QFR! !<7=" RKU RK >'+,?@ FUF !ADS! !<7=" RKU RK >'15A4 FUF !FLEG4CB!b:OPZ!=%dcZ!!#CDF!CW!HTF! F]ALODFS!LOXF!SFNFMCQFS!MOLE!SFWFXHB!ADS!HTFGF! [FGF!DC!SFHFXHAEMF!SOWWFGFDXFB!OD!BJFMFHAM! BHGIXHIGF!EFH[FFD![OMS!H4QF!ADS!LIHADH!FLEG4CB Z!!1TFBF!CEBFGNAHOCDB!ODSOXAHF!HTAH!,EDM"!MCBB! XCLEODFS![OHT!'LQJ!ADShCG!2O]_!SCFB!DCH!MFAS! HC!BJFMFHAM!LAMWCGLAHOCDB!SIGODP!FLEG4CPFDFBOBZ!

PAGE 50

! _>! Discussion +MHTCIPT!XCDBOSFGAEMF!QGCPGFBB!TAB!EFFD!LASF! OD!HTF!QABH!SFXASF!HC!FMIXOSAHF!QAHTCPFDOX! LFXTADOBLB!XAIBODP!ASIMH%CDBFH!',R!HTF!LCMFXI MAG!SFWFXHB!ABBCXOAHFS![OHT!&',!GFLAOD!A! L4BHFG4Z!!1TOB!BHIS4![AB!SFBOPDFS! HC!BQFXOWOXAMM4!HFBH!HTF!T4QCHT FBOB!HTAH!&',!GFBIMHB!WGCL! SFXGFABFS!F]QGFBBOCD!WGC L!HTF!',%MODJFS!PFDFB! +!/0 !ADS! 1234' XCLEODFS![OHT!,3#-! BFYIFBHGAHOCDZ!!$IG!GFBIMHB!QGCNOSF!FNOSFDXF!APAODBH!HTOB!T4QCHTFBOB!EFXAIBF!LOXF!MAXJODP!,EDM"! ADS!'LQJ!ADShCG!2O]_!MONF!WCG!CNFG!A!4FAG!ADS!LI HADH!FLEG4CB!SC!DCH!SOBQMA4!HTF!XTAGAXHFGOBHOX! FLEG4CDOX!SFNFMCQLFDHAM!SFWFXHB!CEBFGNFS!OD!&',! QAHOFDHBZ!!1TOB!BHIS4!OB !HTF!WOGBHR!HC!CIG! JDC[MFSPFR!HC!SOGFXHM4!HFBH!ADS!QGCNOSF!FNOSFDXF !APAODBH!HTF!OSFA!HTAH! XCLEODAHCGOAM!,EDM!MCBB! ADS!'LQJ!ADShCG!2O]_!GFSIXHOCD!XAIBFB!&',Z!!$IG! GFBIMHB!SFLCDBHGAHF!HTAH!AMHFGDAHF!T4QCHTFBFB! DFFS!HC!EF!PFDFGAHFS!ADS!HFBHFS!HC!IDXCNFG!&',!LCMFXIMAG!QAHTCPFDFBOBZ! ',08!ADS!2(e_![FGF!CGOPODAMM4!QGCQCBFS!HC !EF!ODNCMNFS!OD!',"!QA HTCPFDFBOB!EFXAIBF!CW! HTF!QGC]OLOH4!CW!HTFOG!PFDFB!HC !HTF!F]QADSFS!&1)!GFQFAH!HGAXH! ADS!HTF!CEBFGNAHOCD!HTAH!',08! ADS!2(e_!MFNFMB!LA4!EF!XCLQGCLOBFS!OD!',"!HOBBIFB!ADS!XFMMBZ!!9C[FNFGR! +,?@ !ADS! 15A4 JDCXJCIH!LCSFMB!TANF!BIEHMF!QTFDCH4QFB!ADS!WAOM !HC!GFXAQOHIMAHF!LAiCG!',"!XMODOXAM!ADS! TOBHCMCPOXAM!WFAHIGFB!BIXT! AB!BJFMFHAM!LIBXMF!L4CHCDOAR![FAJDFBB!CG![ABHODPR!XAGSOAX! S4BWIDXHOCDR!XCPDOHONF!OLQAOGLFDH!ADS!&',!b"_dcZ!! 1TOB!BIPPFBHB!HTAH!MCBB!CW!',08!CG!2(e_! AMCDF!OB!DCH!BIWWOXOFDH!HC!XAIBF!ASIMH%CDBFH!', !CG!&',R!ADS!HTAH!ASSOHOCDAM!CG!SOWWFGFDH! LCMFXIMAG!SFWFXHB!SOXHAHF!SOBFABFZ!!2ODXF!,3#!BFYIFBHGAHOCD!OB!XMFAGM4!A!QGOLAG4!QAHTCPFDOX! FNFDH!OD!ASIMH%CDBFH!',g!ADS!,3#-R!',08!ADS! 2(e_!LA4!WIDXHOCD!OD!HTF!BALF!CG!QAGAMMFM! QT4BOCMCPOXAM!QAHT[A4BR![F!GFABCDFS!HTAH!XCLE ODAHCGOAM!MCBB!CW!,3#-!ADShCG!',08!LA4! IDSFGMOF!&',Z!!9C[FNFGR!CIG!SAHA!B TC[!HTAH!LOXF!XCDHAODODP!SOBGIQHFS! !<7=R! +,?@ !ADS! 15A4 WAOM!HC!LCSFM!&',Z!!VTAH!AGF!CHTFG!QCBBOEM F!F]QMADAHOCDB!WCG!&',!LCMFXIMAG!SFWFXHBv!

PAGE 51

! _"! &',!LA4!GFBIMH!WGCL!BFYIFBHGAHOCD!CW!AMM!,3#!QGCHFODBZ!!1TF!,3#-!WALOM4!XCDBOBHB!CW! HTGFF!WALOM4!LFLEFGB!b,3#-"R!,3#-=!ADS!,3#KcR!AMM!CW![TOXT!TANF!EFFD!BTC[D!HC!EODS! F]QADSFS!&7)!GFQFAHB!ADS!AGF!F]QGFBBFS!OD!FLEG 4CDOX!LIBXMF!ADS!EGAODR!HTF!H[C!HOBBIFB!HTAH! BFFL!HC!EF!LCBH!AWWFXHFS!OD!&',!b""=R"_`R"_acZ!! 1TFGFWCGFR!AB!&7)!GFQFAHB!PFH!MCDPFG!HTF4! LA4!HOHGAHF!LCGF!,3#-!ADS!OH!OB!DCH!IDHOM!A!HTGFBTCMS!CW!b&1)c u"R>>>! OB!GFAXTFS!EFWCGF!FDCIPT! ,3#-!OB!SFQMFHFS!HC!XAIBF!FLEG4CDOX!SFWFXHB!AD S!&',Z!!.NOSFDXF!APAODBH!HTOB!QCBBOEOMOH4!OB! HTAH! !<7=FUF !LOXFR![TOXT!SFNFMCQ!BFNFGAM!WFAHIGFB!CW !ASIMH%CDBFH!',R!TANF!DC!FLEG4CDOX!CG! XCDPFDOHAM!QTFDCH4QF!b">KcZ!!9C[FNFGR!OH!OB!QCBBO EMF!HTAH!,EDM=!ADShCG! ,EDMK!TANF!CNFGMAQQODP! WIDXHOCDB![OHT!,EDM"!ADS!XCLQFDBAHF!WCG!,EDM "!MCBBZ!!+DCHTFG!AGPILFDH!APAODBH!HTF!,3#-! MCBB%CW%WIDXHOCD!LCSFM!OB!HTAH!',=! GFQFAH!F]QADBOCDB![OHT!b&&1)c u">>> !AMBC!BFYIFBHFG!,3#-! EIH!AGF!DCH!ABBCXOAHFS![OHT!XCDPF DOHAM!SOBFABF!b"_?cZ!!.NOSFDXF! BTC[ODP!HTAH!,3#-!TAB!NAG4ODP! AWWODOHOFB!WCGR!CG!WCGL!SOWWFGFDH!BHGIXHIGFB!CDR!F]QADSFS!b&1)c D !XCLQAGFS!HC!b&&1)c D ![CIMS! QGCNOSF!FNOSFDXF!APAODBH!HTOB!AGPILFDHZ!!#CDFHT FMFBBR!OH![OMM!EF!ODHFGFBHODP!HC!BFF!OW! !<7=R! !<7=L R! !<7=K !SCIEMF!CG!HGOQMF!XCDBHOHIHONF!ADS!HOB BIF%BQFXOWOX!JDCXJCIH!LOXF!SOBQMA4!&',! WFAHIGFBZ!!+MHFGDAHONFM4R!,3#-!"R!=R!ADS!K!MCBB! XCLEODFS![OHT!GFSIXFS!',08!ADShCG!2(e_!LA4! EF!GFYIOGFS!HC!LCSFM!&',Z!!1TOB! T4QCHTFBOB!OB!ODHFGFBHODP!EIH!SOWWO XIMH!HC!HFBH!OD!LOXF!SIF!HC!HTF! XCLQMF]!DAHIGF!CW!HTF!GFYIOGFS!EGFFSODP!BXTFLFZ!! +MHFGDAHONF!AQQGCAXTFB!DFFS!HC!EF!XCDBOSFGFS! HC!SFHFGLODF!OW!ADS!TC[!,3#-!ADS!HTF!',"!MCXIB!XCDHGOEIHF!HC!&',Z!!! +!SOWWFGFDH!LCSFM!HC!F]QMAOD!&',! QAHTCPFDFBOB!QGCQCBFB!HTAH!b&1)c u"R>>>! AMHFG!',"!MCXIB! GFPOCDB!CHTFG!HTADR! CG!OD!ASSOHOCD!HCR! +!/0 !ADS! 1234 Z!!1TF!',"!MCXIB!OB!QAGHOXIMAGM4!ODHFGFBHODP! EFXAIBF!OH!XCDHAODB!HTGFF!PFDFB!XMIBHFGFS!HCPFHT FG!OD!=>!JE!CW!PFDCLOX!'#+!ADS!A!NAGOFH4!CW! GFPIMAHCG4!FMFLFDHB!ADS!ADHOBFDBF!HGADBXGOQHB!TANF !EFFD!LAQQFS!HC!HTF!MCXIBZ!!1TF!',"!MCXIB! XCDBOBHB!CW! +!.+ R! +!/0 !ADS! 1234 Z!!$IG!BHIS4!WCXIBFS!BQFXOWOXAMM4!CD! +!/0 !ADS! 1234'

PAGE 52

! _=! EFXAIBF!HTFGF!AGF!DILFGCIB!GFQCGHB!HTAH! +!/0' ADS! 1234 !MFNFMB!AGF!GFSIXFS!OD!',"!ADS!&',Z!! 9C[FNFGR!HTFGF!AGF!AMBC!GFQCGHB!SFBXGO EODP!',!HOBBIFB!ADS!XFMMB![OHT!GFSIXFS! +!.+ !b"KUcZ!! +!.+ !FDXCSFB!A!HG4QHCQTAD%ABQAGHOX!AXOS!bV'c! GFQFAH!QGCHFOD![OHT!IDJDC[D!WIDXHOCD!ADS!OB! OLLFSOAHFM4!IQBHGFAL!CW! +!/0C'' ',V' OB!BQFXOWOXAMM4!FDGOXTFS!OD!HFBHFB!ADS!HTF!XFDHGAM! DFGNCIB!B4BHFL!b">acZ!!1TFGFWCGFR!OH!OB!QCBBOEMF!HTAH!b&1)c u"R>>>! QCBOHOCDB!DIXMFCBCLFB!OD!A! LADDFG!HTAH!GFSIXFB! +!.+' F]QGFBBOCD!ADS!XCDBFYIFDHM4!GFBIMHB!OD!SFMA4FS!FLEG4CDOX! DFIGCPFDFBOB!ADS!&',Z!!7DWCGHIDAHFM4! +,TJ !LCIBF!JDCXJCIHB!HTAH![CIM S!EF!IBFWIM!OD!HFBHODP! HTOB!T4QCHTFBOB!AGF!DCH!XIGGFDHM4!ANAOMA EMF!ADS![F![FGF!IDAEMF!HC!HAGPFH! +,TJ !OD!LCIBF! FLEG4CDOX!BHFL!XFMMBZ!!! 0GCLCHFG!ADS!ODBIMAHCG!FMFLFDHB!AGF! AMBC!QCBOHOCDFS!QGC]OLAM!HC!HTF!b&1)c D !ADS!AGF! SOWWFGFDHOAMM4!LCSOWOFS!OD!DCGLAM!ADS!',!XFMMB!b"d>cZ!! 1234' QGCLCHFG!FMFLFDHB!ADS!QGCLCHFGB! WCG!ADHOBFDBF!HGADBXGOQHB!AGF!QCBOHOCDF S!OLLFSOAHFM4!SC[DBHGFAL!CW!HTF!b&1)c D !ADS!IQBHGFAL!CW! 1234 Z!!(DBIMAHCGBR! 65E' FMFLFDHB!HTAH!EODS!&1&:!QGCHFODB!A DS!WIDXHOCD!AB!ECIDSAG4!FMFLFDHB!HC! ODTOEOH!TFHFGCXTGCLAHOD!BQGFAS!CG!QGFNFDH!DFAGE4 !QGCLCHFGB!WGCL!AXHONAHODP!ASiAXFDH!PFDFBR!WMADJ! HTF!b&1)c Db"=dc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b&7)c D !DCH!CDM4!EODS!ADS!BFYIFBH FG!,3#-!QGCHFODB!EIH!

PAGE 53

! _K! AMBC!HGOPPFG!08&%LFSOAHFS!T4QFGQTCBQTCG4MAHOC D!ADS!ODXGFABFS!BHAEOMO H4!CW!&7)30"b"d"cZ!! &7)30"!OB!AD!AMHFGDAHONF!BQMOXO DP!WAXHCG!HTAH!WIDXHOCDB!ADHAPCDO BHOXAMM4!HC!,3#-!E4!QGCLCHODP! FLEG4CDOX!AMHFGDAHONF!BQMOXODP!QAHH FGDB!b">UcZ!!1TFGFWCGFR!,3#-!MCBB!XCLEODFS![OHT!ODXGFABFS! &7)30"!MFNFMB!ADShCG!GFSIXFS!',08!ADShCG!2( e_!TANF!HTF!QCHFDHOAM! HC!XCDHGOEIHF!HC!&',! QAHTCMCP4Z!!.NOSFDXF!BIQQCGHODP!A!GCMF!WCG!ODXGFABF S!&7)30"!OD!&',!OB!HTAH!HGADBPFDOX!LOXF! [TOXT!CNFGF]QGFBB!&IPEQ"!SOF!BTCGHM 4!AWHFG!EOGHT!ADS!AGF!GFQCGHFS !HC!TANF!SFMA4FS!L4CPFDFBOB! b"d=cZ!!9C[FNFGR!&7)30"!MFNFMB!AGF!DCGLAMM4! MC[!OD!ASIMHB!ADS!OH! OB!DCH!XMFAG!OW!HTF! B*D>> !TANF!HC]OX!SFMFHFGOCIB!AWWFXHB!OD!ASSOHOCD!HC!CG! ODSFQFDSFDH!CW!,3#-!BFYIFBHGAHOCDR!ODXGFABF S!&7)30"!ADS!QCHFDHOAM!SOBGIQHFS!',"!MCXIB! F]QGFBBOCDZ!!:CG!F]ALQMFR!MCDP!& 7)!GFQFAH!XCDHAODODP!* #+B!LA4!WCGL!BHGIXHIGFB!SOWWFGFDH!WGCL! BTCGHFG!&7)!GFQFAHB![TOXT!QGCNOSF!DCNFM!EODSODP!B OHFB!WCG!ADS!BFYIFBHFG!IDO SFDHOWOFS!WAXHCGBZ!! +MHFGDAHONFM4R!b&1)c u"R>>> !LA4!AXHONAHF!SCIEMF!BHGADSFS!*#+! QAHT[A4BZ!!(DSFFSR!8GCM!ADS! XCMMFPIFB!SFLCDBHGAHFS!HTAH!M CDP!&7)!GFQFAHB!AGF!QGCXFBBFS!ODHC !BLAMM!ODHFGWFG ODP!="!DIXMFCHOSF! *#+B!HTAH!XAD!WIDXHOCD!OD! #&)7E !HC!SC[DGFPIMAHF!b&+)c D !XCDHAODODP!L*#+B!OD!&',! WOEGCEMABHBb"dUcZ!!2LAMM!*#+!E4QGCSIXHB!WGCL!AD HOBFDBF!HGADBXGOQHB!CGOPODAHODP!IQBHGFAL!CW! 1234' TANF!AMBC!EFFD!CEBFGNFS!OD!',!XFMMB!b"d>cZ!! :IGHTFGR!F]QADSFS!&7)!GFQFAH!*#+B!XAD!AXHONAHF! HTF!SCIEMF%BHGADSFS!*#+!bSB*#+c%SFQFDSFDH!QGCHFOD! JODABF!HTAH!DCGLAMM4!WIDXHOCDB!HC!ODTOEOH! HGADBMAHOCD!ADS!QGCLCHF!XFMM!SFAHT! SIGODP!HTF!ODDAHF!OLLIDF!GFBQCDB F!b"d_cZ!!1TFBF!CEBFGNAHOCDB!

PAGE 54

! _U! BIPPFBH!HTAH!&',!ABBCXOAHFS!b&1)c D !LA4!AXHONAHF!SB*#+!QAHT[A4B!AECNF!A!HTGFBTCMS!HTAH! IMHOLAHFM4!SOBGIQHB!ADS!SFMA4B!FLEG4CDOX!SFNFMCQLFDHZ! (D!XCDXMIBOCDR!&',!OB!A!SFEOMOHAHODP!SOBFAB F!HTAH!AWWFXHB!DF[ECGDB!ADS!ASIMHBZ!!1TF! SFNFMCQLFDH!CW!QCHFDHOAM!&',!HT FGAQFIHOXB!GFMOFB!CD!FMIXOSAHODP!HT F!LCMFXIMAG!LFXTADOBLB!HTAH! XAIBF!SOBFABFZ!!$IG!GFQCGH!QGCNOSFB!FNOSFDXF!APAO DBH!A!LCSFM!WCG!XCLEODAHCGOAM!MCBB!CW!,3#-M"! ADShCG!HTF!',%MODJFS!PFDFB! +!/0 !ADS! 1234 Z!!:IGHTFGR![F!SFBXGOEF! AMHFGDAHONF!T4QCHTFBFB!HC! AXXCIDH!WCG!&',Z!!1TFBF!LCSFMB!AGF!DCH!LIHIA MM4!F]XMIBONF!ADS![F!EFMOFNF![AGGADH!WIGHTFG! ODNFBHOPAHOCDZ

PAGE 55

! __! ! ! ! :OPIGF!=%"Z!',"!ADS!',=!AGF!XAIBFS!E4!LOXGCBAH FMMOHF!F]QADBOCD!OD!DCD%XCSODP!GFPOCDB!CG! +!/0 !ADS! 9(:; R!GFBQFXHONFM4Z +c!,IHADH!',08!L*#+!bEM AXJ!MODFc![OHT!HTF!$*:! bPGF4!EC]!MAEFMFS!',08cR!&7)!F]QADBOCDB!OD !HTF!K6%71*!ADS!A!QCM4b+c!HAOM!bw+xD! AGF!ODSOXAHFSZ!!#CHF!HTAH!F]QADBOCDB!XAD! F]XFFS!=R>>>!GFQFAHBZ!!3c!1TF!LIHADH!m#:?! PFDF!bMODFR!ODHGCDBg!CQFD!EC]FBR!71*Bg!XMCB FS!EC]FBR!$*:c![OHT!HTF!&&1)!F]QADBOCD! OD!ODHGCD!"!bF]QADBOCDB!XAD!F]XFFS!""R>>>!GFQFAHBcZ !! !

PAGE 56

! _d! ! ! ! :OPIGF!=%=Z!*#+!'CLODADXF!,CSFMZ!!#CGLAM!AMMF MF!'#+!bEMAXJ!EC]c!GFQFAH B!bPGFFDc!XAD!F]QADS! bGFSc!HC!PFDFGAHF!QAHTCPFDOX!SB*#+!bGFS!TA OGQODc!HTAH!BFYIFBH FGB!,3#-!QGCHFODB! bEMIF!XOGXMFBcZ!!!

PAGE 57

! _`! ! ! ! ! :OPIGF!=%KZ1TGFF!HOPTHM4%MODJFS!PFDFB!AH!HTF! +!!MCXIBZ!!1TF!BHGIXHIGF!CW!HTF!TILAD! +!-' MCXIB! OB!OMMIBHGAHFSZ!!+MBC!BTC[D!AGF!HTF! +!.+ !bGFS!EC]FBcR! +!/0' bCGADPF!EC]FBc!ADS! 1234 !bEMIF!EC]FBc!F]CDB!WC G!AMM!HTGFF!PFDFBZ!!!

PAGE 58

! _a! ! :OPIGF!=%UZ!-AHF!BHAPF!FLEG4CB!MAXJODP!,EDM "!ADS!'LQJ!CG!,EDM"!ADS!2O]_!SC!DCH!BTC[! SFNFMCQLFDHAM!SFWFXHBZ!!+c!*1%0&*! ADAM4BOB!CD!."aZ_!YIASGOXFQB!X'#+Z!! &4XMCQTOMOD!b0QOAc!QGOLFGB![FGF!IBFS!AB!MC ASODP!XCDHGCMZ!!3c!!."`Z_![TCMF!ADS!WO]FS! FLEG4CBZ!!!!

PAGE 59

! _?! ! ! ! :OPIGF!=%_Z!,4CWOEFG!SFNFMCQLFDH!OB!DCH!SFMA4FS!OD! !<7=>+,?@ !CG! !<7=>15A4 !SCIEMF!JDCXJCIHBZ!! VOMS!H4QF!b+!ADS!3cR! !<7=! RKU RK W'+,?@ FUF b&!ADS!'cR!ADS! !<7=! RKU RK W'15A4 FUF b.!ADS! :c!F]HFDBCG!XAGQO!GASOAMOB !HGADBNFGBF!LIBXMFB!CD!."_Z_! bMFWH!QADFMc!ADS!."aZ_!bGOPTH! QADFMc!SFBLOD!bPGFFDc!BHAODFS!HGADBNFGBF !BFXHOCDBZ!!'+0(!bEMIFc!BHAODB!DIXMFOZ!

PAGE 60

! d>! ! ! ! :OPIGF!=%dZ!3CDF!SFWCGLOHOFB!AGF!DCH!CEBFGNFS!OD! !<7=>+,?@ !CG! !<7=>15A4 !SCIEMF!JDCXJCIHBZ!! VOMS!H4QFR! !<7=! RKU RK >+,?@ FUF R!ADS! !<7=! RKU RK >15A4 FUF !BJFMFHCD!QGFQAGAHOCDBZ!! +MXOAD!EMIF!BHAODB!XAGHOMAPF!A DS!AMO\AGOD!GFS!BHAODB!ECDFZ!!! !

PAGE 61

! d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b"dd cZ!!(D!ASSOHOCDR!LO*#+B!XAD!EODS!QGCLCHFG! FMFLFDHB!ADS!ODSIXF!PFDF!F]QGFBBOCD!b"d`R"dacZ!! (H!OB!MOJFM4!HTAH!H TCIBADSB!CW!L*#+B!AGF! S4BGFPIMAHFS!OD!'OXFG!LIHADHBZ!! 1TIBR!LIHAHODP!'OXFG!LA4!TANF!EFFD!HCC!XGISF!AD!AQQGCAXT!HC! FMIXOSAHF!HTF!QGFXOBF!LCMFXIMAG!LFXTADOBLB!E4! [TOXT!LO*#+B!XCDHGOEIHF!HC!FLEG4CDOX!LIBXMF! SFNFMCQLFDHZ!!:IHIGF!BHISOFB!BTCIMS!HAJF!A! LCGF!GFWODFS!AQQGCAXT!ADS!WCXIB!CD!FNAMIAHODP!

PAGE 62

! d=! ODSONOSIAM!CG!BLAMM!PGCIQB!CW!LO*#+B!HC !EFHHFG!IDSFGBHADS!HTFOG!GCMF!OD!LALLAMOAD! SFNFMCQLFDHZ! VF!AMBC!ODNFBHOPAHFS!TC[!MCDP!&1)!F]QADBO CDB!OD!HTF!',08!K6!71*!GFBIMH!OD!&',R!A! SOBFABF!GFBIMHODP!OD!QAGH!WGCL!OLQAOGFS!FLEG 4CDOX!L4CPFDFBOBZ!!$IG!SAHA!SFLCDBHGAHFS!HTAH! GFSIXFS!F]QGFBBOCD!WGCL!HTF!',"%MODJFS!PFDFB! +!/0 !ADS! 1234 !XCLEODFS![OHT!,3#-"!MCBB!OB! DCH!BIWWOXOFDH!HC!LCSFM!&',!OD!LOXFZ!!1TFBF!GFBI MHB!AGPIF!APAODBH!A!GCMF!WCG!',08!ADS!2(e_!OD! &',!QAHTCPFDFBOBZ!!VF!AGF!TCQFWIM!HTAH!CIG!CEB FGNAHOCDB!TOPTMOPTH!HTF!DFXFBBOH4!WCG!WIHIGF! ODNFBHOPAHOCDB!ODHC!HTF!LCMFXIMAG! SFWFXHB!ABBCXOAHFS![OHT!&',Z!!! VTAH!AGF!CHTFG!QCBBOEMF!LFXTADOBLB!HC!F]QMAOD! &',v!!(H!OB!XMFAG!HT AH!,3#-!BFYIFBHGAHOCD! OB!A!QGOLAG4!QAHTCPFDOX!LFXTADOBL!OD!ASIMH% CDBFH!',!ADS!MOJFM4!HTAH!,3#-!MCBB!AMBC! XCDHGOEIHFB!HC!&',Z!!(B!OH!QCBBO EMF!HTAH!BFYIFBHGAHOCD!CW!AMM!,3#-!WALOM4!LFLEFGB!GFBIMHB!OD! &',v!!VF!AGF!XIGGFDHM4!PFDFGAHODP!,3#-"R! ,3#-=!ADS!,3#-K!XCDBHOHIHONF!ADS!XCDSOHOCDAM! LCIBF!JDCXJCIHB!HC!HFBH!HTOB!T4QCHTFBOBZ!! +MHFGDAHONFM4R!&',!LA4!GFBIMH!WGCL!,3#-! BFYIFBHGAHOCD!XCLEODFS![OHT!CH TFG!HC]OX!*#+!FWWFXHBZ!!b&1)c D !GFQFAH!F]QADBOCDB!GFBIMH!OD!HTF! IQGFPIMAHOCD!CW!&7)30"R!AD!,3 #-"!ADHAPCDOBHZ!!1TFGFWCGFR!SO BGIQHODP!HTF!EAMADXF!EFH[FFD! ECHT!,3#-"!ADS!&7)30"!SIGODP!FLEG4CPFDFBOB!L4!GFBIMH!OD!HTF!SFMA4FS!SFNFMCQLFDH! ABBCXOAHFS![OHT!&',Z!!1TOB!T4QCHTFBOB!XCIMS! EF!HFBHFS!E4!PFDFGAHODP!LOXF!HTAH!MAXJ!,3#-! QGCHFODB!ADS!TANF!ODXGFABFS!&7)30"!MFNFMB!OD !BQFXOWOX!HOBBIFB!SIGODP!FLEG4CDOX!SFNFMCQLFDHZ!! 1C]OX!b&7)c D !*#+B!TANF!AMBC!EFFD!BTC[D!HC!EF!QGCXF BBFS!ODHC!BO&7)B!HTAH!XAD!AXH!OD!HGADB!HC! SC[DGFPIMAHF!b&+)c D !XCDHAODODP!HGADBXGOQHBZ!!(H![OMM!EF!ODHFGFBHODP!HC!BFF!OW!LOXF!CNFGF]QGFBBODP! BO&7)B!GFXAQOHIMAHF!&',!WFAHIGFBZ!!-ABHM 4R!OH!OB!WCGLAMM4!QCBBO EMF!HTAH!MCDP!b&1)c D !F]QADBOCDB! TANF!ASSOHOCDAM!HC]OX!QGCQFGHOFB! HTAH!TANF!DCH!EFFD!SO BXCNFGFS!4FHZ!!VF!AGF!TCQFWIM!HTAH!WIHIGF!

PAGE 63

! dK! ODNFBHOPAHOCDB!ODHC!&',![OMM!SFWODF!HTF!LCMFXIMAG!SFWFXHB!HTAH![OMM!FDAEMF!GFBFAGXTFGB!HC!SFNFMCQ! HTFGAQFIHOXB!HC!HGFAH!&',Z!!!!!

PAGE 64

! dU! &9+01.*!U! ,+1.*(+-2!+#'!,.19$'2! Mice Generation and Genotyping 1C!ODAXHONAHF!'OXFG!BQFXOWOXAMM4!OD !BJFMFHAM!LIBXMFR!LOXF!XCDHAODODP!A! +56%& !XCDSOHOCDAM! AMMFMF!b +56%& 6"7J X ![FGF!XGCBBFS!HC!HTCBF!F]QGFBBODP!A! !O"+6&% !HGADBPFDF!b_aRd`cZ!!1TF!GFBIMHODP! +56%& VU6"7J >!O"+6&% !!LAMF!LOXF![FGF!LAHFS!HC! +56%& 6"7JU6"7J !!WFLAMFB!HC!PFDFGAHF! +56%& F U6"7J >!O"+6&% !LIHADH ADOLAMBZ!!2CLF!WFLAMFB!AMBC!XAGGOFS!HTF! IY1PLN' GFQCGHFG!bdacZ!!(H!OB! DCHF[CGHT4!HTAH!LAMFB!XAGG4ODP!A! +56%& 6"7J !AMMFMF!ADS! !O"+6&%' HGADBPFDF!AM[A4B!HGADBLOHHFS!A! +56%& !SFMFHFS!AMMFMF!b +56%&X Z!1TOB!OB!MOJFM4!SIF!HC!MFAJ4! !O"+6&%' F]QGFBBOCD!OD!HTF!LAMF! PFGL-ODFZ!!1TFGFWCGFR!AMM!LIHADH!FLEG4CB![FGF! +56%& FU6"7J g !O"+6&% !ADS!LAMFB![FGF!DFNFG!IBFS! HC!HGADBLOH!HTF! IY1PLN' GFQCGHFGZ! !<7=" RKU RK R! +,?@ FUF !ADS! 15A4 FUF !LOXF!TANF!EFFD!QGFNOCIBM4!SFBXGOEFSZ!!3GOFWM4R! +,?@ FUF LOXF!TANF!'LQJ!F]CDB!"%`!GFQMAXFS![OHT!A! T4PGCL4XOD!GFBOBHADXF!XABBFHHF!b"U"cZ!!! 15A4 FUF ADOLAMB![FGF!FDPODFFGFS!HC!TANF!A! yPAMAXHCBOSABF!GFQCGHFG!PFDF!BIEBHOHIHFS!WCG!2O]_!F]CD"! b"U`cZ!! !<7=" RKU RK !LOXF![FGF!PFDFGAHFS!E4!SFMFHODP!, EDM!F]CD!K![TOXT!FDXCSFB!H[C!*#+! EODSODP!LCHOWB!HTAH!AGF!FBBFDHOAM!WCG!EODSODP!F]QADSF S!&7)!GFQFAH!*#+B!b">KcZ!!+MM!BHGAODB![FGF! LAODHAODFS!CD!A!&_`3-hd5!EAXJPGCIDSZ!!&_`3-hd5 !LOXF![FGF!EGFS!HC!PFDFGAHF![OMS!H4QF! FLEG4CBZ!! !<7=VU RK >'+,?@ FUF LOXF![FGF!XGCBBFS!HC!4OFMS! !<7=" RKU RK >'+,?@ FUF ADOLAMBZ!! !<7=" RKU RK >'15A4 FUF !LOXF![FGF!PFDFGAHFS!E4! !<7=VU RK >'15A4 FUF !XGCBBFB!ADS! !<7=VU RK >'+,?@ VU % g! 15A4 VUF !ODHFGXGCBBFB![FGF!IBFS!HC!PFDFGAHF!,EDM" .Kh .K g!'LQJ lh% g!2O]_ lh% !FLEG4CBZ!!! !!!+MM!LOXF![FGF!PFDCH4QFS!E4!0&*Z!!+SI MH!ADS!FLEG4CDOX!LCIBF!HAOM!BDOQB!bkK!LLc![FGF! M4BFS!OD!_>>!q-!8!EIWWFG!b_>!L,!8& -R!">!L,!1GOB%&MR!Q9!aZ_R!=Z_!L,!,P&M = R!>Z"!LPhL-! PFMAHODR!>ZU_!j!(PFQAMR!>ZU_!j!1[FFD%=>R!">>!qPhL-! 0GCHFODABF!8c!CNFGDOPTH!AH!__z&Z!!1AOM! M4BAHFB![FGF!TFAH!ODAXHONAHFS!E4! ODXIEAHODP!AH!">>z&!WCG!">!LODR!TA OG!QFMMFHFS!E4!XFDHGOWIPAHOCD!AH!

PAGE 65

! d_! LA]OLIL!BQFFS!WCG!_!LOD!OW!DFXFBBAG4!ADS!">!q!IBFS!OD!A!_>!q-!0&*!GFAXHOCD!XCDHAODODP!=Z_!7! 1AY!QCM4LFGABF!b_!0GOLF!0&*!.]HFDSFGcR!"e!HIDODP!EIWWFGR!>ZK`_!L,!S#10B!ADS!HTF!WCMMC[ODP! QGOLFG!BFHB!bK>!QLCM!CW!FAXT!QFG!GFAXHOCDcf! +56%& 6"7J f!'OXFG!:"! b&&1)+&+)1)+&))1&&+++)c!ADS!'OXF G!*"!b&+1)+&1&11&++&1&+++&1cZ!! !O"+6&% f!&GF!:[S!b1)&++&)+)1)+1)+))11&)&++)++&c!ADS!&GF!*FN! b)++&)++&&1))1&)+++1&+)1)&)11cZ!! IY1PLNI f!MAXm!:[S! b11+1&)+1)+)&)1))1))11+1)&c!ADS!MAXm!*FN! b)&)&)1+&+1&)))&+++1++ 1+1&cZ!!,EDM"f!! b&1+&)+1))&1))&1)&++1+1)&&1&+&1)1++)cR! b&)1))&+)+&&&111)+&+&&)++111&c!ADS! b)))11)++1&1&)11+)))+&+&1)))1)1&1)1++cZ!!'LQJf! b)&++)111))))+)&))+1cR!b)++1)+))1&&1)+)&11c!ADS! b))&1))&+&1&1)1&)+1+&&&cZ!! 2O]_f![OMS!H4QF!QGOLFGB! b&&1&1&)&11+&+&+))1&+)&++&c!ADS! b)+&++)&)+&+)+&&+&)&1)11cZ!!2O]_! LIHADH!CMOPCB!b11+1&)+ 1)+)&)1))1))11+1)&c!ADS! b)&)&)1+&+1&)))&+++1++1+1&c!AGF!QCBOHOCDF S!OD!MAXmZ!!)FDCH4QODP!GFAXHOCDB![FGF! 0&*!ALQMOWOFS!HTGCIPT!K>!b,EDM"!ADS!'LQJc!CG!K_! b'OXFGR!&GFR!*=d*!ADS!2O]_c!X4XMFB!CW!?Uz&! WCG!"!LODR!d>z&!WCG!"!LOD!ADS!daz&!WCG!K!LODZ!!! !0&*!WGAPLFDHB![FGF!GF BCMNFS!CD!=Z_j!APAGCBF! PFMBZ!!+MM!F]QFGOLFDHB![FGF!QFGWCGLFS!IDSFG! AQQGCNFS!ADOLAM!QGCHCXCMB!AXXCGSODP!HC!HTF! ODBHOHIHOCDAM!PIOSFMODFB!FBHAEMOBTFS!E4!HT F!7DONFGBOH4!CW!:MCGOSA!(+&7&!XCLLOHHFFZ! RNA Blots and RT-PCR *#+![AB!OBCMAHFS!WGCL!WGFBT! TCLCPFDO\FS!HCDPIFR!FLEG4CDOX! WCGFMOLEB!CG!TODSMOLEB![OHT! 1*(!*FAPFDH!b2OPLAcZ!!:CG!BLAMM!*#+!EMCHBR!AQQG C]OLAHFM4!=>!qP!CW!HCHAM!*#+![AB!GFBCMNFS!CD! ">j!IGFAhQCM4AXG4MALOSF!PFMB! ADS!FMFXHGCEMCHHFS!HC!94ECDS!# l !LFLEGADFB!b+LFGBTALc!AH!=>>!

PAGE 66

! dd! L+!WCG!K!TZ!!3MCHB![FGF!XGCBBMO DJFS!IBODP!A!2HGAHAMODJFG!b2HGAHAPFD c!ADS!QGFT4EGOSO\FS!WCG!AH!MFABH! "!T!AH!K`z&!OD!7-1*+T4E 1, %$MOPC!b+LEOCDc!T4EGOSO\AHOCD!EIWWF G!EFWCGF!CNFGDOPTH!ODXIEAHOCD! AH!K`z&!OD!T4EGOSO\AHOCD!EIWWFG!XCDHAODODP!w K= 0x%FDS%MAEFMFS!QGCEFZ!!0GCEFB![FGF!PFDFGAHFS!E4! FDS%MAEFMODP!=>!QLCMB!CW!'#+!CMOPCDIXMFCHOSF!b( DNOHGCPFDc!XCLQMFLFDHAG4!HC!LO*#+!CG!7d! [OHT!1U!QCM4DIXMFCHOSF!JODABF!b#F[ !.DPMADS!3OCMAEBc!ADS!=_>!q&O!w # % K= 0x!+10!b0FGJOD!.MLFGc! WCMMC[FS!E4!QIGOWOXAHOCD![OHT!,OXGC2QOD 1, !)%=_!XCMILDB!b+LFGBTALcZ!!3MCHB![FGF![ABTFS!b=e! 22&R!>Z"j!2'2c!AH!K`z&!WCG!K>!LOD!WCMMC[FS!E4!H[ C!K>!LOD!GCCL!HFLQFGAHIGF![ABTFBZ!!3MCHB! [FGF!F]QCBFS!HC!8CSAJ!3OC,A]!WOML!ADS!YIAD HOWOXAHOCD![AB!SFHFGLODFS!E4!SFDBOHCLFHG4!CW! AIHCGASOCPGAQTB!IBODP!(LAPF>KZ>=!BCWH[AGFZ!!:CG!*1%0&*R!">! # P!CW!HCHAM!*#+! [AB!GFNFGBF!HGADBXGOEFS!OD!A!=_q-!GFAXHOCD! XCDHAODODP!`_>!DP!GADSCL!TF]ALFGBR!"Z=_qP!bS1c "=% "a R!>Z=!L,!S#10BR!"e!WOGBH BHGADS!EIWWFGR!a!L,!'11R!_>!7!*# ABOD!ADS!=_>!7!2IQFGBXGOQH!(((! GFNFGBF!HGADBXGOQHABF!b(DNOHGCPFDcZ!!0&*!XCDSOHOC DB![FGF!=_z&!WCG!_!LODR!_>z&!WCG!"!TG!ADS!`>z&! WCG!"_!LODZ!!:CMMC[ODP!0&*R!GFAXHOCDB![FGF!HGFAHFS ![OHT!"q-!b=!7c!*#ABF!9!AH!K`z&!WCG!=>!LOD! ADS!=!q-!CW!X'#+![FGF!IBFS!AB!HFLQMAHF!WCG! +56%&' ALQMOWOXAHOCD!OD!A!_>! q-!GFAXHOCD!XCDHAODODP! =Z_7!1GOQMFLABHFG! Q)Z !b.QQFDSCGWcR!"e!EIWWFGR!>ZU!L,!S#10BR!ADS!K>!QLCM!CW!FAXT!CW!HTF! WCMMC[ODP!WCG[AGS!ADS!GFNFGBF!QGOLFGBf! 'OXFGf!b&+&+&)&&1&&1 +&&+&1+&++&+&c!ADS! b))&1)&+1&+1&))+1+)1+&+&&cZ!!,EDM"f! b)11+)1)1&+&+&&++11&)))+&+&c!ADS!b)))&+1&+1))&+11))&1++&cZ! 'LQJf!b)++&++)1)))+&+1)&1)++)+))c !ADS!b&+1&+&))+1))&1)))+&++cZ! 2O]_f!b&1)&&&++)+&&+1&1)))+1)c!A DS!b&1&+)+&1)+)&&1&+1&&+)1&)cZ! 0QOAf!b)&))&+))1&&+1&1+&)c!ADS!b)&&+ 1&&+)&&+11&+)1&1cZ!!0&*!XCDSOHOCDB! [FGF!=`!X4XMFB!CW!?_z&!WCG!K>!BFXR!__z&!WCG!K>!BFX!ADS!`=z&!WCG!"!LOD!ADS!"_!q-!CW!FAXT!0&*! GFAXHOCD![AB!GFBCMNFS!CD!A!">j!AXG4MALOSF!PFM! ADS!F]QCBFS!HC!8CSAJ!3OC,A]!WOMLZ!!!!!!

PAGE 67

! d`! Histology, X-gal Staining, Immunohi stochemistry and TUNEL Assays ,CIBF!FLEG4CB![FGF!XCMMFXHFS!AWHFG!DAHIGAM! CNFGDOPTH!LAHODPB!ADS!TFLAHC]4MOD!ADS!FCBOD! b9n.c!BHAODODP![AB!QFGWCGLFS!E4!BHADSAGS!QG CXFSIGFBZ!!:CG![TCMF! LCIDH!e%PAM!BHAODODPR! FLEG4CB![FGF!TAGNFBHFS!OD!032!ADS!WO]FS!OD! >ZUj!WCGLAMSFT4SF!CNFGDOPTH!AH!Uz&Z!!.LEG4CB! [FGF!GODBFS!HTGFF!HOLFB!WCG!">!LOD!OD !XCDXFDHGAHFS!GODBF!EIWWFG!b>Z"!,!#A9 = 0$ U R!Q9!`ZUR!=!L,! ,P&M = R!>Z=j!(PFQAMR!>Z"j!BCSOIL!SFC]4XTCMAHFc !ADS!BHAODFS![OHT!"!LPhL-!e%PAM!OD! XCDXFDHGAHFS!GODBF!EIWWFGR!_!L,!8 K :Fb&#c d !ADS!_!L,!8 U :Fb&#c d !U%"d!TG!AH!Uz&![OHT!BTAJODPZ!! .LEG4CB![FGF!GODBFS!HTGFF!HOLFB!WCG!_!LOD!OD !032!bQ9!`ZUcR!QCBH%WO]FS!OD!Uj!WCGLAMSFT4SF! CNFGDOPTH!AH!Uz&!ADS!NOBIAMO\FS![OHT!A!-FOXA! SOBBFXHODP!LOXGCBXCQFZ!! :CG!QAGAWWOD!BFXHOCDODPR! FLEG4CB![FGF!WO]FS!OD!Uj!WCGLAMSFT4SF!CNFGDOPTH! AH!Uz&R![ABTFS!OD!032!HTGFF!HOLFB!WCG!">!LOD! ADS!BHCGFS!OD!`>j!.H$9!AH!Uz&Z!!.LEG4CDOX!M OLEB!CG!HCDPIF![FGF!SOBBFXHFS!ADS!SFT4SGAHFS! HTGCIPT!">!LOD![ABTFB!OD!`>j!.H$9R!?_j!.H $9R!">>j!.H$9!ADS!&OHGOBCMN!b:OBTFGcZ!! 'FT4SGAHFS!HOBBIFB![FGF!ODXIEAHF S!OD!QAGAWWOD!HTGFF!HOLFB!WCG!"!TG!IDSFG!NAXIIL!AH!d_z&!EFWCGF! GCIHODF!FLEFSSODPZ!!2FGOAM!BFXHOCDB!b`!qLc![FGF !XIH!CD!A!-FOXA!GCHAG4!LOXGCHCLFR!MAOS!AXGCBB! U_z&![AHFG!HC!GFLCNF![GODJMFBR!QCBOHOCDFS!CD!2IQF GWGCBH!0MIB!LOXGCBXCQF!BMOSFB!b:OBTFGc!ADS! LFMHFS!AH!d_z&!WCG!AH!MFABH!K>!LODZ!!1C!XCLQAGF!BOL OMAG!QMADFB!CW!SOBHAM!WCGFMOLEB!WCG!XCDHGCM!ADS! LIHADH!MOHHFGLAHFB!A!GFWFGFDXF!QCODH![AB!FBHAEMOB TFS!AH!HTF!iIDXHOCD!EFH[FFD!TILFGIBR!GASOIB!ADS! IMDA!ECDFBZ!!1TF!WOGBH!BFXHOCD!HT AH!SOS!DCH!XCDHAOD!AD4! QCGHOCD!CW!TILFGIBR!EIH!BTC[FS!GASOIB!ADS! IMDA![AB!SFFLFS!HTF!GFWFGFDXFZ!!2FXHOCDB!FYIOSO BHADH!WGCL!HTF!GFWFGFDXF!WCG!XCDHGCM!ADS!LIHADH! SOBHAM!WCGFMOLEB![FGF!XCLQAGFS!WCG!AMM!F]QFGOLFDH BZ!!:CG!OLLIDCTOBHCXTFLOBHG4R!BMOSFB![FGF!SF% QAGAWWODO\FS!ADS!GFT4SGAHFS!HTGCIPT!">!LOD![ABT FB!OD!&OHGOBCMNR!">>j!.H$9R!?_j!.H$9R!`>j! .H$9R![AHFG!ADS!032Z!!+DHOPFD!GFHGOFNAM![AB!QFGW CGLFS!WCG!AMM!ADHOECSOFB! E4!ODXIEAHODP!BMOSFB!OD! ">!L,!1GOB!bQ9!?cR!"!L,!.'1+!WCG!K>!LOD!AH!?`z &!ADS!XCCMODP!BMOSFB!OD!HTF!BALF!EIWWFG!AH! GCCL!HFLQFGAHIGF!b*1c!WCG!=>!LODZ!!:CG!LCDCXMCDAM! QGOLAG4!ADHOECSOFBR!BMOS FB![FGF!HGFAHFS![OHT!

PAGE 68

! da! QAQAOD%SOPFBHFS!LCIBF!(P)B!AB!SFBXGOEFS!b"d?cZ!!2 MOSFB![FGF!EMCXJFS!OD!032R!"j!32+R!"j!TFAH! ODAXHONAHFS!PCAH!BFGIL!ADS!>Z>=_j!1[FFD%=>!WCG!"!TG !AH!*1Z!!1TGFF!032!GODBFB![FGF!QFGWCGLFS! ADS!BMOSFB![FGF!HGFAHFS![OHT!QGOLAG4!ADHOECS4! bBFF!EFMC[c!SOMIHFS!OD!EMCXJODP!EIWWFG!AH!Uz&! CNFGDOPTHZ!!2MOSFB![FGF!GODBFS!HTGFF!HOLFB! [OHT!032!ADS!ODXIEAHFS! [OHT!BFXCDSAG4!ADHOECS4! SOMIHFS!OD!EMCXJODP!EIWWFG!WCG!"!TG!AH!*1!OD!HT F!SAGJZ!!1C!GFSIXF!AIHCWM ICGFBXFDXFR!BMOSFB![FGF! ODXIEAHFS![OHT!">!L,!&I2$ U R!_>!L,!&9 K &$$#9 U! bQ9!_Z_c!WCG!K>!LOD!AH!*1![OHT!BTAJODPZ!! 2ALQMFB![FGF!GODBFS!HTGFF!HOLFB!OD!032R!L CIDHFS![OHT!/FXHABTOFMS!XCDHAODODP!'+0(!b/FXHCGc! ADS!NOBIAMO\FS![OHT!A!-FOXA!WMICGFBXFDXF!CG! XCDWCXAM!LOXGCBXCQFZ!! 0GOLAG4!ADHOECSOFB!ADS! SOMIHOCDB![FGF!"f">>!SFBLOD!b'KKR!'+8$cR! "f">>!,9&!b,;K=R!2OPLAc!ADS!"f=>>!XMFANFS! XABQABF%K!b&FMM!2OPDAMODPcZ!17#.-!b*CXTFc![A B!QFGWCGLFS!AXXCGSODP!HC!HTF!LADIWAXHIGFG6B! QGCHCXCMZ!!17#.-!XCLEODFS![OHT!,4C'![AB!QF GWCGLFS!E4!SCODP!,4C'!OLLIDCXTFLOBHG4!WOGBH! AB!SFBXGOEFS!AECNF!WCMMC[FS!E4!17#.-!AXXCGSODPM 4!HC!LADIWAXHIGFG6B!QGCHCXCMZ!!2FXCDSAG4! ADHOECSOFB!IBFS![FGF!"f=_>!PCAH!ADHO%LCIBF!(P )"!XCDiIPAHFS!HC!+MF]A:MICG!Uaa!b(DNOHGCPFDcZ! &MFANFS!XABQABF%K![AB!SFHFXHFS!IBODP!HTF!*AEEOH!(P )!/FXHABHAOD!JOH!OD!XCLEODAHOCD![OHT!/FXHCG! *FS!2IEBHGAHF!b/FXHCG!MAEBcZ!!&C%OLLIDCTOBHCXTF LOBHG4!BQFXOWOX!WCG!XMFANFS!XABQABF%K!ADS! L4CBOD!TFAN4!XTAOD![AB!XCDSIXHFS!E4!XCLEOD ODP!QGOLAG4!ADHOECSOFB!ADS!WCMMC[FS![OHT! XCLEODFS!BFXCDSAG4!ADHOECSOFBZ!!!!!!!!! Immunoblotting 1CDPIFB![FGF!SOBBFXHFS!ADS!QMAXFS!OD!_>% ">>!q-!CW!_>!L,!1GOB{&M!bQ9!s!dZacR!" L,! .'1+R!=j!2'2R!>Z_!L,!QTFD4MLFH T4MBIMWCD4M!WMICGOSFR!_!qPhL! QFQBHAHOD!+R!"!qPhL-! XT4LCBHAHODR!"!L,!ALODCXAQGCOX!AXOSR!" L,! ? %ALODCEFD\ALOSODFR!"!qPhL-!MFIQFQHODR!=!qPhL! AQGCHODOD!ADS!SOBGIQHFS!IBODP!A! LODOAHIGF!QFBHMF!WCMMC[FS!E4!EGOF W!BCDOXAHOCDZ!!.]HGAXHB![FGF! XFDHGOWIPFS!AH!"dR>>>! x D !WCG!"> LODR!ADS!BIQFGDAHADH!QGCHFODB! [FGF!WGAXHOCDAHFS!CD!"=Z_j!2'2% QCM4AXG4MALOSF PFMB!bU>!qP!QGCHFOD!QFG!MADF cZ!!)FMB![FGF!FMFXHGCEMCH HFS!HC!DOHGCXFMMIMCBF!ADS!

PAGE 69

! d?! EMCXJFS!K>!LOD!OD!_j!#CDWAH!SG4!LOMJR!>Z>_j!(P FQAMZ!!3MCHB![FGF!ODXIEAHFS![OHT!QGOLAG4! ADHOECSOFB!bEFMC[c!SOMIHFS!OD!_j!#CDWAH!SG4!LOM JR!>Z>_j!(PFQAM!WCG!"!TG!AH!*1!WCMMC[FS!E4!HTGFF! _!LOD![ABTFB!OD!032R!>Z>_j!(PFQAMZ!!9CGBFGAS OBT!QFGOC]OSABF!XCDiIPAHF S!BFXCDSAG4!ADHOECSOFB! BQFXOWOX!WCG!LCIBF!CG!GAEEOH!(P)!b+LFGBTALc![F GF!SOMIHFS!"f_>>>!OD!_j!#CDWAH!SG4!LOMJR!>Z>_j! (PFQAM!ADS!ODXIEAHFS![OHT!EMCH!WCG!K>!LOD!AH! *1!WCMMC[FS!E4!HTGFF!_!LOD![ABTFB!OD!032R!>Z>_j! (PFQAM!ADS!CDF!_!LOD![ABT!OD!032Z!!(LL IDCEMCHB![FGF!NOBIAMO\FS!E4!.&-!b+LFGBTALcZ!! 2QFXOWOX!QGOLAG4!ADHOECSOFB!ADS!SOMIHOCDB![F GF!"f">>>!ADHO%,4C'!b_Za+R!'AJCcR!"f_>>!ADHO% L4CPFDOD!b:_'R!ABXOHFB!WMIOS!'FNFMCQLFDHAM!2H ISOFB!94EGOSCLAcR!"f">>>!ADHO%L4CBOD!TFAN4! XTAOD!WABH!OBCWCGL!b,;%K=R!2OPLAcR!"f">>>!AD HO%SFBLOD!b'KKR!'AJCc!ADS!"f">R>>>!ADHO%)AQST! bd&_R!+EXALcZ!! Morphometric and Statistical Analysis ,4CWOEFG!DILEFG!ADS!ANFGAPF!XGCBB%BFXHO CDAM!AGFA![FGF!YIADHOWOFS!CD!."dZ_!SFBLOD% BHAODFS!SOBHAM!WCGFMOLE!HGADBNFG BF!BFXHOCDBZ!!(LAPFB![FGF! XAQHIGFS!CD!A!mFOBB!WMICGFBXFDH! LOXGCBXCQF!ADS!L4CWOEFGB![FGF!XCIDHFS!ADS!L FABIGFS!IBODP!+]OCNOBOCD! U!BCWH[AGF!bmFOBBcZ!! 2QFXOWOXAMM4R!A!d>!qL = !EC]![AB!AGEOHGAGOM4!QMAXFS!OD!FAXT!ODSONOSIAM!LIBXMF!LABBZ!!1TF!EC]![AB! BLAMM!FDCIPT!BC!HTAH!AMM!FSPFB![FGF!BIGGCIDSFS!E4 !LIBXMF!ADS!DC!GFPOCD!CW!AD4!EC]!WFMM!CIHBOSF! HTF!LIBXMF!LABBZ!!1TF!DILEFG!CW!L4CWOEFGB![AB! XCIDHFS!OD!FAXT!EC]!ADS!HTF!SAHA!GFQGFBFDHFS!AB! DILEFG!CW!L4CWOEFGB!QFG!XL = !WCG!FAXT!LIBXMFZ!!+!HCHAM!CW!K?a!ADS!=d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

PAGE 70

! `>! LIHADH!MOHHFGLAHFB![FGF!F]ALODFS!AH!FAXT!SFNFMCQL FDHAM!HOLF!QCODHZ!!$DM4!BHAODFS!DIXMFO![OHTOD! LIBXMF!LABBFB![FGF!XCIDHFS!ADS!BHAHOBHOXAM!SO WWFGFDXFB![FGF!SFHFGLODFS!IBODP!2HISFDH6B! # %HFBHZ! Cell Culture, Annexin-V-FITC/Propidium Iodide Staining and Cell Sorting !1C!XGFAHF!HTF! +56%&' XCDSOHOCDAM!,'.*!XFMM!MODFR!LCIBF!FLEG4CDOX!WOEGCEMABHB!SFGONFS! WGCL!FLEG4CB![OHT!H[C! +56%& !XCDSOHOCDAM!AMMFMFB!bBFF!AECNFc![F GF!ODWFXHFS![OHT!A!GFHGCNOGIB! XCDHAODODP!HTF!Q3+3.!NFXHCG! F]QGFBBODP!A!T4PGCL4XOD%GFBOBHADXF!PFDF!ADS!,'.*R!A!,4C'% FBHGCPFD!GFXFQHCG!WIBOCD!QGCHFOD!b9CM MFDEFGP!FH!AMZ!"??KcZ!!1C!LIHAHF! +56%& R!XFMMB![FGF!ODWFXHFS!AH! A!TOPT!,$(![OHT!+S%&,/% 6&% !ASFDCNOGIB!b/FXHCG!3OCMAEBcZ!!&FMMB![FGF!LAODHAODFS!OD!PGC[HT! LFSOIL!b),cR![TOXT!XCDBOBHFS!CW!'IMEFXXC6B! ,CSOWOFS!.APMF!b',.R!)OEXCcR![OHTCIH!QTFDCM! GFSR!XCDHAODODP!">j!WFHAM!ECNODF!BFGIL!b94XM CDFcR!ADS!"j!-%PMIHALODFZ!!Q3+3.%,'.*! ODWFXHFS!XFMMB![FGF!BFMFXHFS!OD!=>>! # PhL-!T4PGCL4XODZ!!1C!ODSIXF!F]QGFBBOCD!CW!,'.*R!XFMMB! [FGF!B[OHXTFS!HC!SOWWFGFDHOAHOCD!LFSOIL!b',c R![TOXT!XCDBOBHFS!CW! ',.!XCDHAODODP!"j!-% PMIHALODFR!>Z_j!TCGBF!BFGILR!">! # PhL-!ODBIMODR!">! # PhL-!HGADBWFGGODR!ADS!"> %` !,! %FBHGASOCMZ!! &FMMB![FGF!PFDCH4QFS!IDSFG!HTF!BALF!XCDSOHOCDB!AB !HTF!LOXF!bBFF!AECNFcZ!!0TABF%XCDHGABH!OLAPFB! CW!IDWO]FSR!IDBHAODFS! +56%&' XCDSOHOCDAM!,'.*!ADS!&GF%ODWFXHFS! +56%&' XCDSOHOCDAM!,'.*!XFMMB! WCMMC[ODP!?d!TCIGB!CW!HGFAHLFDH![OHT!',![OHT!CG![OHTCIH! %FBHGASOCM![FGF!XAQHIGFS!CD!WOML!CD! AD!+]OCNFGH!">!LOXGCBXCQF!bmFOBBcZ!!(LAPFB ![FGF!SOPOHO\FSR!ADS!OW!DFFSFSR!LODCG!MODFAG! ASiIBHLFDHB!HC!XCDHGABH![FGF!LA SF!IBODP!+SCEF!0TCHCBTCQ!BCWH[AGFZ! +56%& XCDSOHOCDAM!,'.*!ADS!&GF%ODWFXHFS! +56%&' XCDSOHOCDAM!,'.*!XFMMB![FGF!XIMHIGFS!OD! HGOQMOXAHF!CD!=U![FMM!HOBBIF%XI MHIGF!HGFAHFS!SOBTFB!OD!',![OHT!CG![OHTCIH! %FBHGASOCM!WCG!=U!TCIGBR! AB!ODSOXAHFSZ!!&FMMB![FGF!HG4QBODO\FS!ADS!XC%BHA ODFS![OHT!+DDF]OD!/%:(1&!ADS!QGCQOSOIL!OCSOSF! IBODP!HTF!+DDF]OD!/%:(1&!+QCQHCBOB!'FHFXHOCD!8 OH!b3OC/OBOCDc!QFG!LADIWAXHIGFG6B!ODBHGIXHOCDBZ!! &FMMB![FGF!BCGHFS!CD!A!:+&2XAD!X4HCLFHFG!b3FXJHCDh'OXJBCDcZ!!+B!A!QCBOHONF!XCDHGCM!WCG!

PAGE 71

! `"! AQCQHCBOB!ADS!HC!SFWODF!PAHODP!QAGALFHFGB R!XFMMB![FGF!HGFAHFS! OD!QAGAMMFM![OHT!_>! # ,! PFMSADAL4XOD!OD!),!WCG!=U!TCIGB!QGOCG!HC!ADAM4B OBR!ADS!FOHTFG!BODPMF%BHAODFS![OHT!+DDF]OD!/% :(1&!CG!QGCQOSOIL!O CSOSFR!CG!SCIEMF%BHAODFSZ! !!+MM!F]QFGOLFDHB![FGF!QFGWCGLFS!IDSFG!A QQGCNFS!ADOLAM!QGCHCXCMB!AXXCGSODP!HC!HTF! ODBHOHIHOCDAM!PIOSFMODFB!FBHAEMOBTFS!E4!HT F!7DONFGBOH4!CW!:MCGOSA!(+&7&!XCLLOHHFFZ! Skeletal preparations ,CIBF!FLEG4CB![FGF!TAGNFBHFS!OD!032R!WO]F S!OD!Uj!WCGLAMSFT4SF!CNFGDOPTH!AH!Uz&R! BJODDFS!ADS!FNOBXFGAHFS!HTF!WCMMC[ODP!SA4Z!!&A GHOMAPF![AB!BHAODFS!E4!ODXIEAHODP!FLEG4CB!OD!>Z=! Ph-!+MXOAD!3MIF!OD!`>j!.H$9hK>! j!AXFHOX!AXOS!CNFGDOPTH!AH!GCCL !HFLQFGAHIGFZ!!1TF!DF]H!SA4! FLEG4CB![FGF!GFT4SGAHFS!E4!ODXIEAHODP!OD! ">>jR!?_jR!`>j!ADS!U>j!.H$9!ADS![AHFG!WCG!K>! LOD!FAXTZ!!#F]HR!ECDF![AB!BHAODFS!E4!GCXJODP!BALQ MFB!OD!>Z"j![hN!+MO\AGOD!*FS!OD!"j!8$9!WCG! H[C!SA4B!AH!GCCL!HFLQFGAHIGFZ!!1TF!+MO\AGOD !*FS!BHAODODP!BCMIHOCD![AB!XTADPFS!CDXF!AWHFG!HTF! WOGBH!=U!TGZ!!2HAODFS!FLEG4CB! [FGF!XMFAGFS!OD!"j!8$9!WCG!IQ! HC!"%U!SA4B!WCMMC[FS!E4!"!TG! ODXIEAHOCDB!FAXT!OD!=_j!PM4XFGCMh`_j!8$9R! _>j!PM4XFGCMh_>j!8$9!ADS!`_j!PM4XFGCMh=_j! 8$9Z!!2HAODFS!ADS!XMFAGFS!FLEG4CB ![FGF!BHCGFS!OD!a>j!PM4XFGCM !ADS!QTCHCPGAQTFS!IBODP!A!-FOXA! BHFGFCBXCQFZ

PAGE 72

! `=! -(21!$:!*.:.*.#&.2! "Z!&GOXJR!:Z9ZR!)GOWWOHTR!5Z2Z!ADS!$GPF MR!-Z.Z!b"?_`c!&CSFB![OHTCIH!&CLLABZ! /&"6'()#='P6)J' 165'['1'P R! 43 R!U"d%U="Z! =Z!,AHHOXJR!5Z2Z!ADS!,AJIDODR!(Z/Z!b=>>dc!#CD%XCSODP!*#+Z! \*,'!"=']%7%# R! 15 Spec No 1 R!*"`%=?Z! KZ!:GOHTR!,Z&ZR!0TFABADHR!,Z!ADS!,AHHOXJR!5Z2Z !b=>>_c!1TF!ALA\ODP!XCLQMF]OH4!CW!HTF!TILAD! HGADBXGOQHCLFZ! R*&'^'\*,']%7%# R! 13 R!a?U%a?`Z! UZ!0TFABADHR!,Z!ADS!,AHHOXJR!5Z2Z!b=>>`c! *AOBODP!HTF!FBHOLAHF!CW!WIDXHOCDAM!TILAD! BFYIFDXFBZ! ]%7",%'I%E R! 17 R!"=U_%"=_KZ! _Z!,AHHOXJR!5Z2Z!ADS!,AJIDODR!(Z/Z!b=>>_c!2LAMM!GFPIMAHCG4!*#+B!OD!LALLAMBZ! \*,'!"=' ]%7%# R! 14 Spec No 1 R!*"="%"K=Z! dZ!3AGHFMR!'Z0Z!b=>>Uc!,OXGC*#+Bf!PFDCLOXB R!EOCPFDFBOBR!LFXTADOBLR!ADS!WIDXHOCDZ! B%== R! 116 R!=a"%=?`Z! `Z!,ABIOR!$Z!ADS!9FAGSR!.Z!b=>>dc!*#+!ADS!QG CHFOD!AXHCGB!OD!e%XTGCLCBCLF!ODAXHONAHOCDZ! B"=J'1?&57D'\)&<'1O,?'_*)7#'`5"= R! 71 R!U"?%U=aZ! aZ!:OMOQC[OX\R!VZR!3TAHHAXTAG44AR!2Z#Z!ADS! 2CDFDEFGPR!#Z!b=>>ac!,FXTADOBLB!CW!QCBH% HGADBXGOQHOCDAM!GFPIMAHOCD!E4!LOXGC*#+Bf!AGF!HTF!ADB[FGB!OD!BOPTHv! ()#'I%8']%7%# R! 9 R! ">=%""UZ! ?Z!-FFR!*Z&ZR!:FODEAILR!*Z-Z!ADS!+LEGCBR!/Z!b "??Kc!1TF!&Z!FMFPADB!TFHFGCXTGCDOX!PFDF!MOD% U!FDXCSFB!BLAMM!*#+B![OHT!ADHOBFDBF!XCLQMFLFDHAGOH4!HC!MOD%"UZ! B%== R! 75 R!aUK%a_UZ! ">Z!VOPTHLADR!3ZR!9AR!(Z!ADS!*INJIDR!)Z!b"??Kc! 0CBHHGADBXGOQHOCDAM!GFPIMAHOCD!CW!HTF! TFHFGCXTGCDOX!PFDF!MOD%"U!E4!MOD%U!LFSOAHFB! HFLQCGAM!QAHHFGD!WCGLAHOCD!OD!&Z!FMFPADBZ! B%== R! 75 R!a__%ad=Z! ""Z!*FODTAGHR!3Z5ZR!2MAXJR!:Z5ZR!3ABBCDR!,ZR!0AB YIODFMMOR!+Z.ZR!3FHHODPFGR!5Z&ZR!*CIPNOFR!+Z.ZR! 9CGNOH\R!9Z*Z!ADS!*INJIDR!)Z!b=>>>c!1T F!="%DIXMFCHOSF!MFH%`!*#+!GFPIMAHFB! SFNFMCQLFDHAM!HOLODP!OD!&AFDCGTAESOHOB!FMFPADBZ! ()#*&% R! 403 R!?>"%?>dZ! "=Z!0ABYIODFMMOR!+Z.ZR!*FODTAGHR! 3Z5ZR!2MAXJR!:ZR!,AGHODSAMFR!,Z >>c!&CDBFGNAHOCD!CW!HTF! BFYIFDXF!ADS!HFLQCGAM!F]QGFBBOCD!CW!MF H%`!TFHFGCXTGCDOX! GFPIMAHCG4!*#+Z! ()#*&% R! 408 R! ad%a?Z! "KZ!-APCB%>Kc!#F[! LOXGC*#+B!WGCL!LCIBF!ADS!TILADZ! I7) R! 9 R!"`_%"`?Z!

PAGE 73

! `K! "UZ!-MANFR!&ZR!eOFR!mZR!8ABBXTAIR!8Z'Z!ADS!&A GGODPHCDR!5Z&Z!b=>>=c!&MFANAPF!CW!2XAGFXGC[% MOJF!L*#+!HAGPFHB!SOGFXHFS!E4!A! XMABB!CW!+GAEOSCQBOB!LO*#+Z! 165%76% R! 297 R!=>_K%=>_dZ! "_Z!*FODTAGHR!3Z5ZR!VFODBHFODR!.Z)ZR!*TCASFBR! ,ZVZR!3AGHFMR!3Z!ADS!3AGHFMR!'Z0Z!b=>>=c! ,OXGC*#+B!OD!QMADHBZ! ]%7%E'+%8 R! 16 R!"d"d%"d=dZ! "dZ!+GANODR!+Z+ZR!-APCB%>Kc!1TF!BLAMM!*#+!QGCWOMF!SIGODP! 'GCBCQTOMA!LFMADCPABHFG!SFNFMCQLFDHZ! +%8'B%== R! 5 R!KK`%K_>Z! "`Z!8OLR!5ZR!8GOXTFNBJ4R!+ZR!)GASR!;ZR!9A4FBR!)Z'ZR!8CBOJR!8Z2ZR!&TIGXTR!)Z,Z!ADS!*INJIDR! )Z!b=>>Uc!(SFDHOWOXAHOCD!CW! LAD4!LOXGC*#+B!HTAH!XCQIGOW 4![OHT!QCM4GOECBCLFB!OD! LALLAMOAD!DFIGCDBZ! /&"6'()#='P6)J'165'['1'P R! 101 R!Kd>%Kd_Z! "aZ!-OLR!-Z0ZR!)MABDFGR!,Z.ZR!;FJHAR!2ZR!3IGP FR!&Z3Z!ADS!3AGHFMR!'Z0Z!b=>>Kc!/FGHFEGAHF! LOXGC*#+!PFDFBZ! 165%76% R! 299 R!"_U>Z! "?Z!-OLR!-Z0ZR!-AIR!#Z&ZR!VFODBHFODR!.Z)ZR!+ ESFMTAJOLR!+ZR!;FJHAR!2ZR!*TCASFBR!,ZVZR! 3IGPFR!&Z3Z!ADS!3AGHFMR!'Z0Z!b=>>Kc!1T F!LOXGC*#+B!CW!&AFDCGTAESOHOB!FMFPADBZ! ]%7%E' +%8 R! 17 R!??"%">>aZ! =>Z!3FGF\OJCNR!.ZR!)IG4FNR!/ZR!NAD!SF!3FMHR!5ZR! VOFDTCMSBR!.ZR!0MABHFGJR!*Z9Z!ADS!&IQQFDR!.Z! b=>>_c!0T4MCPFDFHOX!BTASC[ODP!ADS!XCLQIHA HOCDAM!OSFDHOWOXAHOCD! CW!TILAD!LOXGC*#+! PFDFBZ! B%== R! 120 R!="%=UZ! ="Z!-F[OBR!3Z0ZR!3IGPFR!&Z3Z!ADS!3AGHFMR!'Z0Z !b=>>_c!&CDBFGNFS!BFFS! QAOGODPR!CWHFD!WMADJFS! E4!ASFDCBODFBR!ODSOXAHFB!HT AH!HTCIBADSB!CW!TILAD!PF DFB!AGF!LOXGC*#+!HAGPFHBZ! B%== R! 120 R! "_%=>Z! ==Z!-FFR!;ZR!5FCDR!8ZR!-FFR!5Z1ZR!8OLR!2Z! ADS!8OLR!/Z#Z!b=>>=c! ,OXGC*#+!LAHIGAHOCDf! BHFQ[OBF!QGCXFBBODP!ADS! BIEXFMMIMAG!MCXAMO\AHOCDZ! R,<"'^ R! 21 R!UddK%Ud`>Z! =KZ!-AIR!#Z&ZR!-OLR!-Z0ZR!VFODBHFODR!.Z)Z!ADS!3A GHFMR!'Z0Z!b=>>"c!+D!AEIDSADH!XMABB!CW!HOD4! *#+B![OHT!QGCEAEMF!GFPIMAHCG4!GC MFB!OD!&AFDCGTAESOHOB!FMFPADBZ! 165%76% R! 294 R!a_a%ad=Z! =UZ!&AOR!eZR!9APFSCGDR!&Z9Z!ADS!&IMMFDR!3Z*Z! b=>>Uc!9ILAD!LOXGC*#+B !AGF!QGCXFBBFS!WGCL! XAQQFSR!QCM4ASFD4MAHFS!HGADBXGOQHB!HT AH!XAD!AMBC!WIDXHOCD!AB!L*#+BZ! I7) R! 10 R!"?_`%"?ddZ! =_Z!-FFR!;ZR!8OLR!,ZR!9ADR!5ZR!;FCLR!8Z9ZR!FFR!2ZR!3AFJR!2Z9Z!ADS!8OLR!/Z#Z!b=>>Uc! ,OXGC*#+!PFDFB!AGF!HGADBXGO EFS!E4!*#+!QCM4LFGABF!((Z! R,<"'^ R! 23 R!U>_"%U>d>Z! =dZ!;OR!*ZR!>Kc!.]QCGHOD%_!LFSOAHFB!HTF!DIXMFAG! F]QCGH!CW!QGF%LOXGC*#+B!ADS!BTCGH!TAOGQOD!*#+BZ! ]%7%E'+%8 R! 17 R!K>""%K>"dZ!

PAGE 74

! `U! =`Z!-FFR!;ZR!+TDR!&ZR!9ADR!5ZR!&TCOR!9ZR!8OLR!5ZR!;OLR!5ZR!-FFR!5ZR!0GCNCBHR!0ZR!*ASLAGJR!$ZR! 8OLR!2Z '%#')=C !b=>>Kc!1TF!DIXMFAG!*#ABF!(((!'GCB TA!ODOHOAHFB!LOXGC*#+!QGCXFBBODPZ! ()#*&% R! 425 R!U"_%U"?Z! =aZ!-IDSR!.ZR!)IHHODPFGR!2ZR!&AMASCR!+ZR!'ATMEFGPR! 5Z.Z!ADS!8IHA4R!7Z!b=>>Uc!#IXMFAG!F]QCGH! CW!LOXGC*#+!QGFXIGBCGBZ! 165%76% R! 303 R!?_%?aZ! =?Z!&IMMFDR!3Z*Z!b=>>Uc!1GADBXGOQHOCD!ADS!QG CXFBBODP!CW!TILAD!LOXGC*#+!QGFXIGBCGBZ! !"=' B%== R! 16 R!ad"%ad_Z! K>Z!9ALLCDSR!2Z,Z!b=>>_c!'OXODP!ADS!BMOXODP!1TF! XCGF!LAXTODFG4!CW!HTF!*#+!ODHFGWFGFDXF! QAHT[A4Z! :R`1'a%## R! 579 R!_a==%_a=?Z! K"Z!9IHNAPDFGR!)Z!ADS!mALCGFR!0Z'Z!b=>>=c !+!LOXGC*#+!OD!A!LIMHOQMF%HIGDCNFG!*#+O! FD\4LF!XCLQMF]Z! 165%76% R! 297 R!=>_d%=>d>Z! K=Z!mFDPR!;ZR!VAPDFGR!.Z5Z!ADS!&IMMFDR!3Z*Z!b =>>=c!3CHT!DAHIGAM!ADS!SFBOPDFS!LOXGC!*#+B! XAD!ODTOEOH!HTF!F]QGFBBOCD!CW!XCPDAHF! L*#+B![TFD!F]QGFBBFS!OD!TILAD!XFMMBZ! !"='B%== R! 9 R!"K=`%"KKKZ! KKZ!mFDPR!;ZR!;OR!*Z!ADS!&IMMFDR!3Z*Z!b=>>Kc!,O XGC*#+B!ADS!BLAMM!OD HFGWFGODP!*#+B!XAD! ODTOEOH!L*#+!F]QGFBBOCD!E4!BOLOMAG!LFXTADOBLBZ! /&"6'()#='P6)J'165'['1'P R! 100 R!?``?% ?`aUZ! KUZ!'CFDXTR!5Z)ZR!0FHFGBFDR!&Z0Z!ADS!2TAGQR!0Z +Z!b=>>Kc!BO*#+B!XAD!WIDXHOCD!AB!LO*#+BZ! ]%7%E'+%8 R! 17 R!UKa%UU=Z! K_Z!3APPAR!2ZR!3GAXTHR!5ZR!9IDHFGR!2ZR!,ABBOGFGR!8ZR!9CMH\R!5ZR!.AXTIBR!*Z!ADS!0ABYIODFMMOR!+Z.Z! b=>>_c!*FPIMAHOCD!E4!MFH%`!ADS! MOD%U!LO*#+B!GFBIMHB!OD! HAGPFH!L*#+!SFPGASAHOCDZ! B%== R! 122 R!__K%_dKZ! KdZ!-OIR!5ZR!&AGLFMMR!,Z+ZR!*ONABR!:Z/ZR! ,AGBSFDR!&Z)ZR!1TCLBCDR!5Z,ZR!2CDPR!5Z5ZR! 9ALLCDSR!2Z,ZR!5CBTIA%1CGR!-Z!ADS!9ADDCDR!)Z 5Z!b=>>Uc!+GPCDAIHF=!OB!HTF!XAHAM4HOX! FDPODF!CW!LALLAMOAD!*#+OZ! 165%76% R! 305 R!"UK`%"UU"Z! K`Z!.MEABTOGR!2Z,ZR!-FDSFXJFMR!VZ!ADS!1IBXTMR! 1Z!b=>>"c!*#+!ODHFGWFGFDXF!OB!LFSOAHFS!E4! ="%!ADS!==%DIXMFCHOSF!*#+BZ! ]%7%E'+%8 R! 15 R!"aa%=>>Z! KaZ!0OMMAOR!*Z2ZR!3TAHHAXTAG44AR!2Z#ZR!+GHI BR!&Z)ZR!mCMMFGR!1ZR!&CIPCHR!#ZR!3AB4IJR!.ZR! 3FGHGADSR!.Z!ADS!:OMOQC[OX\R!VZ!b=>>_c!(DTO EOHOCD!CW!HGADBMAHOCDAM!ODOHOAHOCD!E4!-FH%`! ,OXGC*#+!OD!TILAD!XFMMBZ! 165%76% R! 309 R!"_`K%"_`dZ! K?Z!0OMMAOR!*Z2ZR!3TAHHAXTAG44AR!2Z#Z!ADS!: OMOQC[OX\R!VZ!b=>>`c!*FQGFBBOCD!CW!QGCHFOD! B4DHTFBOB!E4!LO*#+Bf!TC[!LAD4!LFXTADOBLBv! Q&%7JE'B%=='`5"= R! 17 R!""a%"=dZ!

PAGE 75

! `_! U>Z!.DGOPTHR!+Z5ZR!5CTDR!3ZR!)AIMR!7ZR!1IBXTMR!1ZR!2ADSFGR!&Z!ADS!,AGJBR!'Z2Z!b=>>Kc! ,OXGC*#+!HAGPFHB!OD!'GCBCQTOMAZ! ]%7",%'`5"= R! 5 R!*"Z! U"Z!-F[OBR!3Z0ZR!2TOTR!(Z9ZR!5CDFB%*TCASFBR! ,ZVZR!3AGHFMR!'Z0Z!ADS!3IGPFR!&Z3Z!b=>>Kc! 0GFSOXHOCD!CW!LALLAMOAD!LOXGC*#+!HAGPFHBZ! B%== R! 115 R!`a`%`?aZ! U=Z!2HAGJR!+ZR!3GFDDFXJFR!5ZR!*IBBFMMR!*Z3Z! ADS!&CTFDR!2Z,Z!b=>>Kc!(SFDHOWOXAHOCD!CW! 'GCBCQTOMA!,OXGC*#+!HAGPFHBZ! /a"1'`5"= R! 1 R!.d>Z! UKZ!5CTDR!3ZR!.DGOPTHR!+Z5ZR!+GANODR!+ZR!1IBXTMR!1 ZR!2ADSFGR!&Z!ADS!,AGJBR!'Z2Z!b=>>Uc!9ILAD! ,OXGC*#+!HAGPFHBZ! /a"1'`5"= R! 2 R!FKdKZ! UUZ!8OGOAJOSCIR!,ZR!#FMBCDR!0Z1ZR!8CIGADCNR!+ZR! :OH\OFNR!0ZR!3CI4OCIJCBR!&ZR!,CIGFMAHCBR!mZ! ADS!9AH\OPFCGPOCIR!+Z!b=>>Uc!+!XCLEODFS!XCL QIHAHOCDAM%F]QFGOLFDHAM !AQQGCAXT!QGFSOXHB! TILAD!LOXGC*#+!HAGPFHBZ! ]%7%E'+%8 R! 18 R!""d_%""`aZ! U_Z!)GIDR!'ZR!VADPR!;Z-ZR!-ADPFDEFGPFGR!'ZR!) IDBAMIBR!8Z&Z!ADS!*AiF[BJ4R!#Z!b=>>_c! LOXGC*#+!HAGPFH!QGFSOXHOCDB!AXGCBB!BFNF D!'GCBCQTOMA!BQFXOFB!ADS!XCLQAGOBCD!HC! LALLAMOAD!HAGPFHBZ! /a"1'B",?*#'`5"= R! 1 R!F"KZ! UdZ!8GFJR!+ZR!)GIDR!'ZR!0C4R!,Z#ZR!VCMWR!*ZR!*C BFDEFGPR!-ZR!.QBHFODR!.Z5ZR!,AX,FDALODR!0ZR! SA!0OFSASFR!(ZR!)IDBAMIBR!8Z&ZR!2HCWWFMR!,Z '%#')=C !b=>>_c!&CLEODAHCGOAM!LOXGC*#+!HAGPFH! QGFSOXHOCDBZ! ()#']%7%# R! 37 R!U?_%_>>Z! U`Z!eOFR!eZR!-IR!5ZR!8IMECJABR!.Z5ZR!)CMIER!1Z *ZR!,CCHTAR!/ZR!-ODSEMAS%1CTR!8ZR!-ADSFGR!.Z2Z! ADS!8FMMOBR!,Z!b=>>_c!24BHFLAHOX!SOBXCNFG4! CW!GFPIMAHCG4!LCHOWB!OD!TILAD!QGCLCHFGB!ADS! Kt!71*B!E4!XCLQAGOBCD!CW!BFNFGAM!LALLAMBZ! ()#*&% R! 434 R!KKa%KU_Z! UaZ!VOFDTCMSBR!.ZR!8MCCBHFGLADR!VZ0ZR!,OBJAR! .ZR!+MNAGF\%2AANFSGAR!.ZR!3FGF\OJCNR!.ZR!SF! 3GIOiDR!.ZR!9CGNOH\R!9Z*ZR!8AIQQODFDR!2Z!ADS! 0MABHFGJR!*Z9Z!b=>>_c !,OXGC*#+!F]QGFBBOCD! OD!\FEGAWOBT!FLEG4CDOX!SFNFMCQLFDHZ! 165%76% R! 309 R!K">%K""Z! U?Z!2FLQFGFR!-Z:ZR!:GFFLADHMFR!2ZR!0OHTA%*C[FR!(ZR!,CBBR!.ZR!'LOHGCNBJ4R!.Z!ADS!+LEGCBR!/Z! b=>>Uc!.]QGFBBOCD!QGCWOMODP!CW!LALLAMOAD! LOXGC*#+B!IDXCNFGB!A!BIEBFH!CW!EGAOD% F]QGFBBFS!LOXGC*#+B![OHT!QCBBOEMF!GCMFB!OD !LIGODF!ADS!TILAD!DFIGCDAM!SOWWFGFDHOAHOCDZ! ]%7",%'`5"= R! 5 R!*"KZ! _>Z!mTACR!;ZR!*ADBCLR!5Z:ZR!-OR!+ZR!/FSADHTALR! /ZR!NCD!'GFTMFR!,ZR!,IHTR!+Z#ZR!1BIXTOTABTOR! 1ZR!,X,ADIBR!,Z1ZR!2XT[AGH\R!*Z5Z!ADS! 2GONABHANAR!'Z!b=>>`c !'4BGFPIMAHOCD!CW! XAGSOCPFDFBOBR!XAGSOAX!XCDSIXHOCDR!ADS!XF MM!X4XMF!OD!LOXF!MAXJODP!LO*#+%"%=Z! B%== R! 129 R! K>K%K"`Z! _"Z!-OIR!#ZR!3F\QGC\NADDA4AR!2ZR!VOMMOALBR!+Z9 ZR!>ac!LOXGC*#+%"KKA!GFPIM AHFB!XAGSOCL4CX4HF!QGCMOWFGAHOCD!ADS! BIQQGFBBFB!BLCCHT!LIBXMF!PFDF !F]QGFBBOCD!OD!HTF!TFAGHZ! ]%7%E'+%8 R! 22 R!K=U=%K=_UZ!

PAGE 76

! `d! _=Z!NAD!*CCOiR!.ZR!2IHTFGMADSR!-Z3ZR!-OIR!#ZR! VOMMOALBR!+Z9ZR!,X+DAMM4R!5ZR!)FGAGSR!*Z'ZR! *OXTAGSBCDR!5Z+Z!ADS!$MBCDR!.Z#Z!b=>>dc!+! BOPDAHIGF!QAHHFGD!CW!BHGFBB%GFBQCDBONF! LOXGC*#+B!HTAH!XAD!FNCJF!XAGSO AX!T4QFGHGCQT4!ADS!TFAGH!WAOMIGFZ! /&"6'()#='P6)J'165'['1' P R! 103 R!"a=__%"a=d>Z! _KZ!VADPR!2ZR!+IGCGAR!+Z3ZR!5CTDBCDR!3Z+ZR!>ac! 1TF!FDSCHTFMOAM%BQFXOWOX!LOXGC*#+!LO*%"=d! PCNFGDB!NABXIMAG!ODHFPG OH4!ADS!ADPOCPFDFBOBZ! +%8'B%== R! 15 R!=d"%=`"Z! _UZ!3AFJR!'ZR!/OMMFDR!5ZR!2TODR!&ZR!&ALAGPCR!:Z'ZR!)4POR!2Z0Z!ADS!3AGHFMR!'Z0Z!b=>>ac!1TF! OLQAXH!CW!LOXGC*#+B!CD!QGCHFOD!CIHQIHZ! ()#*&% R! 455 R!dU%`"Z! __Z!2FMEAXTR!,ZR!2XT[ADTAIBBFGR!3ZR!1TOFGWFMSF GR!#ZR!:ADPR!mZR!8TADODR!*Z!ADS!*AiF[BJ4R!#Z! b=>>ac!VOSFBQGFAS!XTADPFB!OD!QGCHFO D!B4DHTFBOB!ODSIXFS!E4!LOXGC*#+BZ! ()#*&% R! 455 R! _a%dKZ! _dZ!3FGDBHFODR!.ZR!8OLR!2Z;ZR!&AGLFMMR!,Z+ZR!,IGXTOBCDR!.Z0ZR!+MXCGDR!9ZR!-OR!,ZmZR!,OMMBR! +Z+ZR!.MMFSPFR!2Z5ZR!+DSFGBCDR!8Z/Z!ADS!9ADDCDR! )Z5Z!b=>>Kc!'OXFG!OB!FBBFDHOAM!WCG!LCIBF! SFNFMCQLFDHZ! ()#']%7%# R! 35 R!="_%="`Z! _`Z!,IGXTOBCDR!.Z0ZR!0AGHGOSPFR!5Z:ZR!1ALR! $Z9ZR!&TFMCIWOR!2Z!ADS!9ADDCDR!)Z5Z!b=>>_c! &TAGAXHFGO\AHOCD!CW!'OXFG%SFWOXOF DH!LIGODF!FLEG4CDOX!BHFL!XFMMBZ! /&"6'()#='P6)J'165'['1' P R! 102 R!"="K_%"="U>Z! _aZ!9AGWFR!3Z'ZR!,X,ADIBR!,Z1ZR!,ADBWOFMSR!5Z9 ZR!9CGDBHFODR!.Z!ADS!1A EODR!&Z5Z!b=>>_c!1TF! *#ABF(((!FD\4LF!'OXFG!OB!GFYIOGFS!WCG!LCGQTCPFDFBOB!EIH!DCH!QAHHFGDODP!CW!HTF!NFGHFEGAHF! MOLEZ! /&"6'()#='P6)J'165'['1'P R! 102 R!">a?a%">?>KZ! _?Z!+DSMR!1ZR!,IGXTOBCDR!.Z0ZR!-OIR!:ZR!mTADPR!; ZR!;IDHA%)CD\AMF\R!,ZR!1CEOABR!5ZVZR!+DSMR! &Z'ZR!2F4JCGAR!5Z1ZR!9ADDCDR!)Z5Z!ADS!,OMM AGR!2Z.Z!b=>>dc!1TF!LO*#+%QGCXFBBODP! FD\4LF!SOXFG!OB!FBBFDHOAM!WCG!HTF!LCGQTCPF DFBOB!ADS!LAODHFDADXF!CW!TAOG!WCMMOXMFBZ! B*&&' `5"= R! 16 R!">U"%">U?Z! d>Z!'ALOADOR!'ZR!+MF]ADSFGR!5Z5ZR!$t*CIGJF R!5Z*ZR!,X,ADIBR!,ZR!5ASTANR!+Z0ZR!&FQJCR!&Z-ZR! 9AIB[OGHTR!VZVZR!9AGWFR!3Z'Z! ADS!2HGFHHCOR!.Z!b=>>ac!'O XFG!ODAXHONAHOCD!MFASB!HC! QGCPGFBBONF!WIDXHOCDAM!A DS!BHGIXHIGAM!SFPFDFGAHOCD!CW!HTF!LCIBF!GFHODAZ! ^'(%*&"E65 R! 28 R! Ua`a%Uaa`Z! d"Z!9A4ABTOR!8ZR!&TINA!SF!2CIBA!-CQFBR!2Z, ZR!8ADFSAR!,ZR!1ADPR!:ZR!9AiJCNAR!0ZR!-ACR!8ZR! $t&AGGCMMR!'ZR!'ABR!0Z0ZR!1AGAJTCNBJ4R!+ZR!,OBJAR!.Z+Z '%#')=C !b=>>ac!,OXGC*#+! EOCPFDFBOB!OB!GFYIOGFS!WCG!LCIBF!QGOLCGSOAM! PFGL!XFMM!SFNFMCQLFDH!ADS!BQFGLAHCPFDFBOBZ! /a"1'Y(R R! 3 R!F"`KaZ! d=Z!9AGGOBR!8Z2ZR!mTADPR!mZR!,X,ADIBR!,Z1ZR!9AGW FR!3Z'Z!ADS!2IDR!eZ!b=>>dc!'OXFG!WIDXHOCD! OB!FBBFDHOAM!WCG!MIDP!FQOHTFMOIL!LCGQTCPFDFBOBZ! /&"6'()#='P6)J'165'['1'P R! 103 R!==>a% =="KZ!

PAGE 77

! ``! dKZ!,IGXTOBCDR!.Z0ZR!2HFODR!0ZR!eIADR!mZR!0A DR!9ZR!mTADPR!,Z>`c!&GOHOXAM!GCMFB!WCG!'O XFG!OD!HTF!WFLAMF!PFGLMODFZ! ]%7%E'+%8 R! 21 R!da=%d?KZ! dUZ!3FGSDOJR!'ZR!:ADR!+Z0ZR!0CHHFGR!&Z5Z!ADS! -ICR!-Z!b=>>ac!,OXGC*#+!QGCXFBBODP!QAHT[A4! GFPIMAHFB!CMWAXHCG4!DF IGCD!LCGQTCPFDFBOBZ! B*&&'`5"= R! 18 R!"`_U%"`_?Z! d_Z!&IFMMAGR!1Z-ZR!'ANOBR!1Z9ZR!#FMBCDR!0Z1ZR!-CFER!)Z3ZR!9AGWFR!3Z'ZR!7MMOADR!.Z!ADS! ,X,ADIBR!,Z1Z!b=>>ac!'OXFG!MCBB!OD!BHGO AHAM!DFIGCDB!QGCSIXFB!EFTANOCGAM!ADS! DFIGCADAHCLOXAM!QTFDCH4QFB!OD!HT F!AEBFDXF!CW!DFIGCSFPFDFGAHOCDZ! /&"6'()#='P6)J'165'[' 1'P R! 105 R!_d"U%_d"?Z! ddZ!'ANOBR!1Z9ZR!&IFMMAGR!1Z-ZR!8CXTR!2Z,ZR!3A GJFGR!+Z5ZR!9AGWFR!3Z'ZR!,X,ADIBR!,Z1Z!ADS! 7MMOADR!.Z,Z!b=>>ac!&CDSOHOCDAM!MCBB!CW!'OXFG !SOBGIQHB!XFMMIMAG!ADS!HOBBIF!LCGQTCPFDFBOB! OD!HTF!XCGHF]!ADS!TOQQCXALQIBZ! ^'(%*&"E65 R! 28 R!UK==%UKK>Z! d`Z!&TFDR!5Z&ZR!,CGHOLFGR!5ZR!,AGMF4R!5Z!ADS!)C MSTALFGR!'Z5Z!b=>>_c!,4C'%XGF!HGADBPFDOX! LOXFf!A!LCSFM!WCG!XCDSOHOCDAM!LIHAPFDFBOB!A DS!MODFAPF!HGAXODP!CW!BJFMFHAM!LIBXMFZ! ]%7%E5E R! 41 R!""d%"="Z! daZ!2CGOADCR!0Z!b"???c!)FDFGAMO\FS!MAXm!F]QG FBBOCD![OHT!HTF!*$2+=d!&GF!GFQCGHFG!BHGAODZ! ()#' ]%7%# R! 21 R!`>%`"Z! d?Z!:IJAPA[AR!1ZR!#CPALOR!,ZR!;CBTOJA[AR!,ZR!(JFDCR!,ZR!$JA\AJOR!1ZR!1AJALOR!;ZR! #AJA4ALAR!1Z!ADS!$BTOLIGAR!,Z!b=>>Uc!'O XFG!OB!FBBFDHOAM!WCG!WCGLAHOCD!CW!HTF! TFHFGCXTGCLAHOD!BHGIXHIGF!OD!NFGHFEGAHF!XFMMBZ! ()#'B%=='`5"= R! 6 R!`aU%`?"Z! `>Z!9CMMFDEFGPR!2Z,ZR!&TFDPR!0Z:Z!ADS!VFODHG AIER!9Z!b"??Kc!7BF!CW!A!XCDSOHOCDAM!,4C'! HGADBXGOQHOCD!WAXHCG!OD!BHISOFB!CW!,4C'! HGADB%AXHONAHOCD!ADS!LIBXMF!SFHFGLODAHOCDZ! /&"6%%J57DE'"S'#$%'()#5"7)='P6)J%,O'"S'16 5%76%E'"S'#$%'[75#%J'1#)#%E'"S'P,%&56) R! 90 R! a>=a%a>K=Z! `"Z!3FGPBHGCLR!'Z+ZR!0FDDR!3Z9ZR!2HGADSR!+ZR!0FGG 4R!*Z-ZR!*ISDOXJOR!,Z+Z!ADS!1AQBXCHHR!2Z5Z! b=>>=c!0GCLCHFG%BQFXOWOX!GFPIM AHOCD!CW!,4C'!EODSODP!ADS!BO PDAM!HGADBSIXHOCD!XCCQFGAHF! HC!QAHHFGD!PFDF!F]QGFBBOCDZ! !"=%6*=)&'6%== R! 9 R!_a`%d>>Z! `=Z!*CBFDEFGPR!,Z(ZR!)FCGPFBR!2Z+ZR!+BA[AXTAO XTAGDR!+ZR!+DAMAIR!.Z!ADS!1AQBXCHHR!2Z5Z! b=>>dc!,4C'!ODTOEOHB!:BHM"!ADS!7HGD!F]QGFB BOCD!E4!ODSIXODP!HGADBX GOQHOCD!CW!LO*%=>dZ! Q$%'^"*&7)='"S'6%=='<5"="DO R! 175 R!``%a_Z! `KZ!mTACR!;ZR!2ALAMR!.Z!ADS!2GONABHANAR!'Z!b =>>_c!2FGIL!GFBQCDBF!WAXHCG!GFPIMAHFB!A!LIBXMF% BQFXOWOX!LOXGC*#+!HTAH!HAGPFHB! 9ADS=!SIGODP!XAGSOCPFDFBOBZ! ()#*&% R! 436 R!="U%==>Z! `UZ!$t*CIGJFR!5Z*ZR!2[ADBCDR!,Z2Z!ADS!9A GWFR!3Z'Z!b=>>dc!,OXGC*#+B!OD!LALLAMOAD! SFNFMCQLFDH!ADS!HILCGOPFDFBOBZ! `5&#$'+%S%6#E'I%E'B'R,<&O"'Q"J)O R! 78 R!"`=%"`?Z!

PAGE 78

! `a! `_Z!9CGDBHFODR!.ZR!,ADBWOFMSR!5Z9ZR!;FJHAR! 2ZR!9IR!5Z8ZR!9AGWFR!3Z'ZR!,X,ADIBR!,Z1ZR! 3ABJFGNOMMFR!2ZR!3AGHFMR!'Z0Z!ADS!1AEO DR!&Z5Z!b=>>_c!1TF!LOXGC*#+!LO*%"?d!AXHB! IQBHGFAL!CW!9C]Ea!ADS!2TT!OD!MOLE!SFNFMCQLFDHZ! ()#*&% R! 438 R!d`"%d`UZ! `dZ!-FFR!&Z1ZR!*OBCLR!1Z!ADS!2HGAIBBR!VZ,Z!b=>>dc!,OXGC*#+B!OD!LALLAMOAD!SFNFMCQLFDHZ! `5&#$'+%S%6#E'I%E'B'R,<&O"'Q"J)O R! 78 R!"=?%"K?Z! ``Z!2CJCMR!#Z2Z!ADS!+LEGCBR!/Z!b=>>_c!,FBCSFGL AMM4!F]QGFBBFS!'GCBCQTOMA!LOXGC*#+%"!OB! GFPIMAHFS!E4!1[OBH!ADS!OB!GFYIOGFS! OD!LIBXMFB!SIGODP! MAGNAM!PGC[HTZ! ]%7%E'+%8 R! 19 R!=KUK% =K_UZ! `aZ!8[CDR!&ZR!9ADR!mZR!$MBCDR!.Z#Z!ADS!2GONABHANA R!'Z!b=>>_c!,OXGC*#+"!ODWMIFDXFB!XAGSOAX! SOWWFGFDHOAHOCD!OD!'GCBCQTOMA! ADS!GFPIMAHFB!#CHXT!BOPDAMODPZ! /&"6'()#='P6)J'165'['1'P R! 102 R!"a?ad%"a??"Z! `?Z!-ICR!'ZR!*FDAIMHR!/Z,Z!ADS!*ADSCR!1Z+Z!b =>>_c!1TF!GFPIMAHOCD!CW!#CHXT!BOPDAMODP!OD! LIBXMF!BHFL!XFMM!AXHONAHOCD!ADS!QCBHDAHAM!L4CPFDFBOBZ! 1%,57'B%=='+%8'`5"= R! 16 R!d"=%d==Z! a>Z!*ACR!0Z8ZR!8ILAGR!*Z,ZR!:AGJTCDSFTR!,ZR!3A BJFGNOMMFR!2Z!ADS!-CSOBTR!9Z:Z!b=>>dc! ,4CPFDOX!WAXHCGB!HTAH!GFPIMAHF!F]QGF BBOCD!CW!LIBXMF%BQFXOWOX!LOXGC*#+BZ! /&"6'()#=' P6)J'165'['1'P R! 103 R!a`="%a`=dZ! a"Z!3FDF\GAR!*ZR!'ANOBR!*Z-ZR!-CXJBTCDR!'ZR! 1IGDFGR!'Z-Z!ADS!VFODHGAIER!9Z!b"??>c!1TF! QGCHFOD!(Sf!A!DFPAHONF!GFPIMAHCG!CW! TFMO]%MCCQ%TFMO]!'#+!EODSODP!QGCHFODBZ! B%== R! 61 R!U?% _?Z! a=Z!-IR!5ZR!VFEER!*ZR!*OXTAGSBCDR!5Z+Z!ADS!$M BCDR!.Z#Z!b"???c!,4C*f!A!LIBXMF%GFBHGOXHFS! EABOX!TFMO]%MCCQ%TFMO]!HGADBXGO QHOCD!WAXHCG!HTAH!ADHAPCDO \FB!HTF!AXHOCDB!CW!,4C'Z! /&"6' ()#='P6)J'165'['1'P R! 96 R!__=%__`Z! aKZ!8OLR!9Z8ZR!-FFR!;Z2ZR!2ONAQGABASR!7ZR!,AM TCHGAR!+Z!ADS!'IHHAR!+Z! b=>>dc!,IBXMF%BQFXOWOX! LOXGC*#+!LO*%=>d!QGCLCHFB!LIBXMF!SOWWFGFDHOAHOCDZ! ^'B%=='`5"= R! 174 R!d``%da`Z! aUZ!9IQQFGH\R!3Z!ADS!9FGGMFGR!+Z!b=>>_c!*FPIMA HOCD!CW!QGCMOWFGAHOCD!ADS!AQCQHCBOB!SIGODP! SFNFMCQLFDH!CW!HTF!QGFOLQMADH AHOCD!FLEG4C!ADS!HTF!QMAXFDHAZ! `5&#$'+%S%6#E'I%E'B' R,<&O"'Q"J)O R! 75 R!=U?%=d"Z! a_Z!&TADR!5Z+ZR!8GOXTFNBJ4R!+Z,Z!ADS!8CBOJR!8Z 2Z!b=>>_c!,OXGC*#+%="!OB!AD!ADHOAQCQHCHOX! WAXHCG!OD!TILAD!PMOCEMABHCLA!XFMMBZ! B)76%&'I%E R! 65 R!d>=?%d>KKZ! adZ!&TFDR!5Z:ZR!,ADSFMR!.Z,ZR!1TCLBCDR!5Z,ZR!VIR!>dc!1TF!GCMF!CW! LOXGC*#+%"!ADS!LOXGC*#+%"KK!OD!BJFMFHAM! LIBXMF!QGCMOWFGAHOCD!ADS!SOWWFGFDHOAHOCDZ! ()#']%7%# R! 38 R!==a%=KKZ! a`Z!0GCNCBHR!0ZR!2OMNFGBHFODR!*Z+ZR!'OBTAGHR!' ZR!VAMWGOSBBCDR!5ZR!'iIQFSAMR!(ZR!8DOCMAR!3ZR! VGOPTHR!+ZR!2ALIFMBBCDR!3ZR!*ASLAGJR!$Z!ADS! .J[AMMR!8Z!b=>>=c!'OXFG!OB!GFYIOGFS!WCG!

PAGE 79

! `?! XTGCLCBCLF!BFPGFPAHOCD!ADS!PFDF! BOMFDXODP!OD!WOBBOCD!4FABH!XFMMBZ! /&"6'()#='P6)J'165'['1' P R! 99 R!"ddUa%"dd_KZ! aaZ!9AMMR!(Z,ZR!#CLAR!8Z!ADS!)GF[AMR!2Z(Z! b=>>Kc!*#+!ODHFGWFGFDXF!LAXTODFG4!GFPIMAHFB! XTGCLCBCLF!S4DALOXB!SIGODP!LOHCBOB!ADS!LFOCBOB!OD!WOBBOCD!4FABHZ! /&"6'()#='P6)J'165'[' 1'P R! 100 R!"?K%"?aZ! a?Z!&AMODR!)Z+Z!ADS!&GCXFR!&Z,Z!b=>>dc!,OXG C*#+%XADXFG!XCDDFXHOCDf!HTF!EFPODDODP!CW!A! DF[!HAMFZ! B)76%&'I%E R! 66 R!`K?>%`K?UZ! ?>Z!9AGQFGR!0Z2Z!b"?a?c! !O"#"756'+OE#&"?$O Z!=!FSZ!2AIDSFGBR!-CDSCDZ! ?"Z!&TCR!'Z9Z!ADS!1AQBXCHHR!2Z5Z!b=>>`c!, 4CHCDOX!S4BHGCQT4f!FLFG PODP!LFXTADOBLB!WCG! ',"!ADS!',=Z! `5"6$5,'`5"?$OE'P6#) R! 1772 R!"?_%=>UZ! ?=Z!+BMADOSOBR!&ZR!5ADBFDR!)ZR!+LFLO4AR!&ZR!2TI HMFGR!)ZR!,ATASFNADR!,ZR!1BOMWOSOBR!&ZR!&TFDR! &ZR!+MMFLADR!5ZR!VCGLBJALQR!#Z)ZR!/CCOiBR!,Z '%#')=C !b"??=c!&MCDODP!CW !HTF!FBBFDHOAM! L4CHCDOX!S4BHGCQT4!GFPOCD!ADS!LAQQODP!CW!HTF!QIHAHONF!SFWFXHZ! ()#*&% R! 355 R!_Ua%__"Z! ?KZ!3GCCJR!5Z'ZR!,X&IGGAXTR!,Z.ZR!9AGMF4R!9Z)ZR!3I XJMFGR!+Z5ZR!&TIGXTR!'ZR!+EIGAHADOR!9ZR! 9IDHFGR!8ZR!2HADHCDR!/Z0ZR!1TOGOCDR!5Z0ZR!9ISBCDR!1Z '%#')=C !b"??=c!,CMFXIMAG!EABOB!CW! L4CHCDOX!S4BHGCQT4f!F]QADBOCD! CW!A!HGODIXMFCHOSF!b&1)c!GF QFAH!AH!HTF!Kt!FDS!CW!A! HGADBXGOQH!FDXCSODP!A!QGCHFO D!JODABF!WALOM4!LFLEFGZ! B%== R! 69 R!Ka_Z! ?UZ!:IR!;Z9ZR!0O\\IHOR!+ZR!:FD[OXJR!*Z)ZR!5GZR!8ODPR!5ZR!*AiDAGA4ADR!2ZR!'IDDFR!0ZVZR!'IEFMR! 5ZR!#ABBFGR!)Z+ZR!+BTO\A[AR!1ZR!SF!5CDPR!0Z '%#')=C !b"??=c!+D!IDBHAEMF!HGOQMFH!GFQFAH!OD!A! PFDF!GFMAHFS!HC!L4CHCDOX!LIBXIMAG!S4BHGCQT4Z! 165%76% R! 255 R!"=_d%"=_aZ! ?_Z!,ATASFNADR!,Z2ZR!+LFLO4AR!&ZR!5ADBFDR! )ZR!2AECIGODR!-ZR!3AOGSR!2ZR!#FNOMMFR!&Z.ZR! VCGLBJALQR!#ZR!2FPFGBR!3ZR!3AH\FGR!,ZR!-ALFGSODR!5Z '%#')=C !b"??Kc!2HGIXHIG F!ADS!PFDCLOX! BFYIFDXF!CW!HTF!L4CHCDOX!S4BHGCQT4!b',!JODABFc!PFDFZ! \*,'!"=']%7%# R! 2 R!=??%K>UZ! ?dZ!-OYICGOR!&Z-ZR!(JFSAR!;ZR!VFAHTFGBQCCDR!,ZR! *OXJFGR!8ZR!2XTCBFGR!3Z)ZR!'AMHCDR!5Z&ZR!'A4R! 5ZVZ!ADS!*ADILR!-Z0Z!b=>>Kc!,4CHCDOX!S4B HGCQT4!H4QF!=f!TILAD! WCIDSFG!TAQMCH4QF!ADS! FNCMIHOCDAG4!XCDBFGNAHOC D!CW!HTF!GFQFAH!HGAXHZ! P,'^'\*,']%7%# R! 73 R!aU?%ad=Z! ?`Z!,ADJCSOR!+ZR!-ODR!eZR!3MA]AMMR!3Z&ZR!2[ADB CDR!,Z2Z!ADS!1TCGDHCDR!&Z+Z!b=>>_c!#IXMFAG! *#+!WCXO!OD!HTF!TFAGH!OD!L4CHCDOX!S4BHGCQT4Z! B5&6'I%E R! 97 R!""_=%""__Z! ?aZ!,ADJCSOR!+ZR!1FDP%7LDIA4R!0ZR!8G4LR!,ZR!9F DSFGBCDR!'ZR!2[ADBCDR!,Z!ADS!1TCGDHCDR! &Z+Z!b=>>Kc!*OECDIXMFAG!ODXMIBOCDB!OD!BJFMFH AM!LIBXMF!OD!L4CHCDOX!S4BHGCQT4!H4QFB!"!ADS! =Z! P77'(%*&"= R! 54 R!`d>%`daZ! ??Z!,ADJCSOR!+ZR!7GEODAHOR!&Z*ZR!;IADR!>"c!,IBXMFEMODS! MCXAMO\FB!HC!DIXMFAG!WCXO!CW!AEFGGADH !*#+!OD!L4CHCDOX!S4BHGCQT4!H4QFB!"!ADS!=Z! \*,'!"=' ]%7%# R! 10 R!="d_%="`>Z!

PAGE 80

! a>! ">>Z!,OMMFGR!5ZVZR!7GEODAHOR!&Z*ZR!1FDP%7LDIA4R!0 ZR!2HFDEFGPR!,Z)ZR!34GDFR!3Z5ZR!1TCGDHCDR! &Z+Z!ADS!2[ADBCDR!,Z2Z!b=>>>c!*FXGIOHLFDH!CW !TILAD!LIBXMFEMODS!QGCHFODB!HC!b&7)cbDc! F]QADBOCDB!ABBCXOAHFS![OHT!L4CHCDOX!S4BHGCQT4Z! R!`Y'^ R! 19 R!UUK?%UUUaZ! ">"Z!VTFFMFGR!1Z,ZR!8G4LR!,Z&Z!ADS!1TCGDHCDR! &Z+Z!b=>>`c!*OECDIXMFAG!WCXO!AH!HTF! DFIGCLIBXIMAG!iIDXHOCD!OD !L4CHCDOX!S4BHGCQT4!H4QF!"Z! (%*&",*E6*='+5E"&J R! 17 R!=U=%=U`Z! ">=Z!;IADR!;ZR!&CLQHCDR!2Z+ZR!2CEX\AJR!8ZR!2HF DEFGPR!,Z)ZR!1TCGDHCDR!&Z+ZR!)GOWWOHTR!5Z'Z! ADS!2[ADBCDR!,Z2Z!b=>>`c!,IBXMFEMODS%MOJF!"!OD HFGAXHB![OHT!*#+!TAOGQODB!OD!BQMOXODP! HAGPFH!ADS!QAHTCPFDOX!*#+BZ! (*6=%56'P65JE'I%E R! 35 R!_U`U%_UadZ! ">KZ!8ADASOAR!*Z#ZR!5CTDBHCDFR!8Z+ZR!,ADJCSO R!+ZR!-IDPIR!&ZR!1TCGDHCDR!&Z+ZR!.BBCDR!'ZR! 1OLLFGBR!+Z,ZR!9AIB[OGHTR!VZVZ!ADS!2[AD BCDR!,Z2Z!b=>>Kc!+!LIBXMFEMODS!JDCXJCIH! LCSFM!WCG!L4CHCDOX!S4BHGCQT4Z! 165%76% R! 302 R!"?`a%"?a>Z! ">UZ!9CR!1Z9ZR!&TAGMFHR!3Z#ZR!0CIMCBR!,Z)ZR! 2ODPTR!)ZR!2[ADBCDR!,Z2Z!ADS!&CCQFGR!1Z+Z! b=>>Uc!,IBXMFEMODS!QGCHFODB!GF PIMAHF!AMHFGD AHONF!BQMOXODPZ! R!`Y'^ R! 23 R!K">K%K""=Z! ">_Z!2ANJIGR!*Z2ZR!0TOMOQBR!+Z/Z!ADS!&CCQFGR!1Z +Z!b=>>"c!+EFGGADH!GFPIMAHOCD!CW!ODBIMOD! GFXFQHCG!AMHFGDAHONF!BQMOXODP!OB!ABBCXOAHFS! [OHT!ODBIMOD!GFBOBHADXF!OD!L4CHCDOX!S4BHGCQT4Z! ()#']%7%# R! 29 R!U>%U`Z! ">dZ!0TOMOQBR!+Z/ZR!1OLXTFDJCR!-Z1Z!ADS!&CCQFGR! 1Z+Z!b"??ac!'OBGIQHOCD!CW!BQMOXODP!GFPIMAHFS! E4!A!&7)%EODSODP!QGCHFOD!OD!L4CHCDOX!S4BHGCQT4Z! 165%76% R! 280 R!`K`%`U"Z! ">`Z!:ODBHFGFGR!5Z!b=>>=c! ,4CHCDOX!S4BHGCQT4!H4QF!=Z! R*&'^'(%*&"= R! 9 R!UU"%UU`Z! ">aZ!VFBHFGMAJFDR!5Z9ZR!/AD!SFG!mFFR!&Z.ZR!0F HFGBR!VZ!ADS!VOFGODPAR!3Z!b=>>Kc!1TF!',V'! QGCHFOD!WGCL!HTF!L4CHCDOX!S4BHGCQT4!b',"c!PF DF!GFPOCD!OB!SFNFMCQLFDHAMM4!GFPIMAHFS! ADS!OB!QGFBFDH!LCBH!QGCLODFDHM4 !OD!B4DAQBF%SFDBF!EGAOD!AGFABZ! `&)57'I%E R! 971 R!""d%"=`Z! ">?Z!VFBHFGMAJFDR!5Z!b=>>Kc!'OBBFGHAHOCDR! 7DONFGOBOH4!CW!#OiLFPFDR!#OiLFPFDZ! "">Z!2LOHTR!1Z!b"???c!1TF!V'!GFQFAHf!A!XC LLCD!AGXTOHFXHIGF!WCG!OSNFGBF!WIDXHOCDBZ! Q&%7JE' `5"6$%,'165 Z! """Z!2AGJAGR!0Z2ZR!9ADR!5Z!ADS!*FSS4R!2Z!b=>>Uc!(D !BOHI!T4EGOSO\AHOCD!AD AM4BOB!CW!'LQJ!L*#+! OD!ASIMH!LCIBF!HOBBIFBZ! (%*&",*E6*='+5E"&J R! 14 R!U?`%_>dZ! ""=Z!8ADASOAR!*Z#ZR!7GEODAHOR!&Z*ZR!&GIBBFMMFR!/Z5 ZR!-ICR!'ZR!-FFR!;Z5ZR!9AGGOBCDR!5Z8ZR!$TR!2Z0Z! ADS!2[ADBCDR!,Z2Z!b=>>Kc!'FNFMCQLFDHAM!F]QGFBBOCD!CW!LCIBF!LIBXMFEMODS!PFDFB! ,EDM"R!,EDM=!ADS!,EDMKZ! ]%7%'RA?&'/)##%&7E R! 3 R!U_?%Ud=Z! ""KZ!1OLXTFDJCR!-ZR!#ABHAODX\4JR!VZR!2XTDFOSFGR!1ZR! 0AHFMR!3ZR!9CWLADDR!:Z!ADS!&ABJF4R!&Z1Z! b"??_c!:IMM%MFDPHT!L4CHCDOD!QGCH FOD!JODABF!b`=!J'Ac!SOBQMA 4B!BFGODF!JODABF!AXHONOH4Z! /&"6' ()#='P6)J'165'['1'P R! 92 R!_Kdd%_K`>Z!

PAGE 81

! a"! ""UZ!,IGAD4OR!+ZR!mTADPR!*ZR!-OIR!:ZR!9OGADCR!8ZR!(H CR!,ZR!.QBHFODR!9Z:Z!ADS!9AGHBTCGDFR!'Z5Z! b=>>"c!,4CHCDOX!S4BHGCQT4!QGCHFOD!JODABF! QTCBQTCG4MAHFB!HTF!L4CBOD!QTCBQTAHABF! HAGPFHODP!BIEIDOH!ADS! ODTOEOHB!L4CBOD!QTCBQTAHABF!AXHONOH4Z! :R`1'a%## R! 493 R!a>%aUZ! ""_Z!8AMOLADR!0ZR!&AHAMIXXOR!'ZR!-ALR!5Z1ZR!8 CDSCR!*ZR!)IHOFGGF\R!5Z&ZR!*FSS4R!2ZR!0AMAXODR!,ZR! mCG\ADCR!+ZR!&TOFDR!8Z*Z!ADS!*IO\%-C\ADCR!0Z !b=>>_c!,4CHCDOX!S4BHGCQT4!QGCHFOD!JODABF! QTCBQTCG4MAHFB!QTCBQTCMALEAD!ADS!GFPIMAHFB!XAMXOIL!IQHAJF!OD!XAGSOCL4CX4HF! BAGXCQMABLOX!GFHOXIMILZ! ^'`5"='B$%, R! 280 R!a>"d%a>="Z! ""dZ!VADBODJR!'Z)ZR!NAD!9FGQFDR!*Z.ZR!&CFG [ODJFM%'GOFBBFDR!,Z,ZR!)GCFDFDR!0Z5ZR! 9FLLODPBR!3Z+Z!ADS!VOFGODPAR!3Z!b=>>Kc! +MHFGDAHONF!BQMOXODP! XCDHGCMB!L4CHCDOX! S4BHGCQT4!QGCHFOD!JODABF!BHGIXH IGFR!FD\4LAHOX!AXHONOH4R!A DS!BIEXFMMIMAG!MCXAMO\AHOCDZ! !"=' B%=='`5"= R! 23 R!_Ua?%__>"Z! ""`Z!,IBBODOR!(ZR!3OGAMR!'ZR!,AGO DR!$ZR!:IGMADR!2Z!ADS!2AMNAHCGOR!2Z!b"???c!,4CHCDOX!S4BHGCQT4! QGCHFOD!JODABF!F]QGFBBFS!OD!GAH!XAGSOAX!LIBXMF !OB!ABBCXOAHFS![OHT!BAGXCQMABLOX!GFHOXIMIL! ADS!PAQ!iIDXHOCDBZ! ^'\5E#"6$%,'BO#"6$%, R! 47 R!KaK%K?=Z! ""aZ!,IGAJALOR!;ZR!$THCR!9ZR!(JFSAR!7ZR!2TOLAS AR!8ZR!,CLCOR!1Z!ADS!8A[AJALOR!8Z!b"??ac! 0GCLCHFG!CW!L',+90h2O]_f!SOWWFGFDHOAM!IHOM O\AHOCD!CW!LIMHOQMF!HGADBXGOQHOCD!ODOHOAHOCD! BOHFB!ADS!QCBOHONFhDFPAHONF!GFPIMAHCG4!FMFLFDHBZ! \*,'!"=']%7%# R! 7 R!=">K%=""=Z! ""?Z!0TALR!;Z&ZR!,ADR!#ZR!9CMHR!(ZR!2F[G4R!&Z+ZR!0AMMR!)ZR!5CTDBCDR!8Z!ADS!,CGGOBR!)Z.Z!b=>>_c! &TAGAXHFGOBAHOCD!CW!HTF!HGADBXGOQHOCD!WAXH CGR!2(e_R!IBODP!A!DF[!QADFM!CW!LCDCXMCDAM! ADHOECSOFBZ! ^'B%=='`5"6$%, R! 95 R!??>%">>"Z! "=>Z!9AGGOBR!2Z.ZR!VODXTFBHFGR!&Z-Z!ADS!5CTDB CDR!8Z5Z!b=>>>c!:IDXHOCDAM!ADAM4BOB!CW!HTF! TCLFCSCLAOD!QGCHFOD!2(e_Z! (*6=%56'P65JE'I%E R! 28 R!"a`"%"a`aZ! "="Z!8A[AJALOR!8ZR!$THCR!9ZR!1AJO\A[AR!1Z!ADS! 2AOHCR!1Z!b"??dc!(SFDHOWOXAHOCD!ADS!F]QGFBBOCD! CW!BO]!WALOM4!PFDFB!OD!LCIBF!GFHODAZ! :R`1'a%## R! 393 R!=_?%=dKZ! "==Z!2QOH\R!:ZR!'FLOPDCDR!5ZR!0CGHFIR!+ZR!8ATDR!+ZR!&CDX CGSFHR!5Z0ZR!'AFPFMFDR!'Z!ADS!,AOGFR!0Z! b"??ac!.]QGFBBOCD!CW!L4CPFDOD!SIGODP!FLEG4CPFDF BOB!OB!XCDHGCMMFS!E4!2O]hBODF!CXIMOB! TCLFCQGCHFODB!HTGCIPT!A!XCDB FGNFS!,.:K!EODSODP!BOHFZ! /&"6'()#='P6)J'165'['1'P R! 95 R! "U==>%"U==_Z! "=KZ!2AHCR!2ZR!#AJALIGAR!,ZR!&TCR!'Z9ZR!1AQBXCHHR! 2Z5ZR!$\AJOR!9Z!ADS!8A[AJALOR!8Z!b=>>=c! (SFDHOWOXAHOCD!CW!HGADBXGOQHOCDAM!HAGPFHB!WC G!2O]_f!OLQMOXAHOCD!WCG!HTF!QAHTCPFDFBOB!CW! L4CHCDOX!S4BHGCQT4!H4QF!"Z! \*,'!"=']%7%# R! 11 R!">U_%">_aZ! "=UZ!VADPR!;Z9ZR!+LOGTAFGOR!2ZR!8ADPR!2ZR!VFMMBR!*Z 'Z!ADS!)GOWWOHTR!5Z'Z!b"??Uc!0GFWFGFDHOAM! DIXMFCBCLF!ABBFLEM4!AH!'#+!HGOQMFH!GF QFAHB!WGCL!HTF!L4CHCDOX!S4BHGCQT4!PFDFZ! 165%76% R! 265 R!dd?%d`"Z!

PAGE 82

! a=! "=_Z!$HHFDR!+Z'Z!ADS!1AQBXCHHR!2Z5Z!b"??_c!1G OQMFH!GFQFAH!F]QADBOC D!OD!L4CHCDOX!S4BHGCQT4! AMHFGB!HTF!ASiAXFDH!XTGCLAHOD!BHGIXHIGFZ! /&"6'()#='P6)J'165'['1'P R! 92 R!_Ud_%_Ud?Z! "=dZ!:OMOQQCNAR!)Z#ZR!1TOFDFBR!&Z0ZR!0FDDR!3Z9ZR! &TCR!'Z9ZR!9IR!;Z5ZR!,CCGFR!5Z,ZR!8MFBFGHR! 1Z*ZR!-CEADFDJCNR!/Z/Z!ADS!1AQBXCHHR!2Z5Z! b=>>"c!&1&:%EODSODP!BOHFB!WMADJ!&1)h&+)! GFQFAHB!ADS!WCGL!A!LFHT4MAHOCD%BFDBOHONF!ODBIMAHCG!AH!HTF!',"!MCXIBZ! ()#']%7%# R! 28 R!KK_% KUKZ! "=`Z!:OMOQQCNAR!)Z#Z!b=>>ac!)FDFHOXB!ADS!FQOPFD FHOXB!CW!HTF!LIMHOWIDX HOCDAM!QGCHFOD!&1&:Z! B*&&'Q"?'+%8'`5"= R! 80 R!KK`%Kd>Z! "=aZ!3TAPANAHOR!2ZR!)TAHQADSFR!+Z!ADS!-FIDPR!3Z!b"??dc!#CGLAM!MFNFMB!CW!',!*#+!ADS! L4CHCDOD!QGCHFOD!JODABF!OD!BJ FMFHAM!LIBXMF!WGCL!ASIMH!L 4CHCDOX!S4BHGCQT4!b',c!QAHOFDHBZ! `5"6$5,'`5"?$OE'P6#) R! 1317 R!"__%"_`Z! "=?Z!(DIJAOR!+ZR!'C4IR!,ZR!8AHCR!1ZR!-OADPR!;ZR!8I GIR!2ZR!;ALALCHCR!,ZR!8CEA4ABTOR!;Z!ADS! 2CEIFR!)Z!b=>>>c!*FSIXFS!F]QGFBBOCD!CW !',+90h2(e_!PFDF!OD!L4CHCDOX!S4BHGCQT4! LIBXMFZ! !*E6=%'(%&8% R! 23 R!"U="%"U=dZ! "K>Z!2AECIGOR!-Z+ZR!,ATASFNADR!,Z2ZR!#AGADPR!,ZR!-FFR!'Z2ZR!2IGTR!-Z&Z!ADS!8CGDFMIJR!*Z)Z! b"??Kc!.WWFXH!CW!HTF!L4CHCDOX!S4BHGCQT4!b' ,c!LIHAHOCD!CD!L*#+!MFNFMB!CW!HTF!',! PFDFZ! ()#']%7%# R! 4 R!=KK%=KaZ! "K"Z!#CNFMMOR!)ZR!)FDDAGFMMOR!,ZR!mFMADCR!)ZR!0O \\IHOR!+ZR!:AHHCGODOR!&ZR!&ABJF4R!&Z1Z!ADS! 'AMMAQOXXCMAR!3Z!b"??Kc!:AOMIGF!OD!SFHFXHODP!L* #+!HGADBXGOQHB!WGCL!HTF!LIHAHFS!AMMFMF!OD! L4CHCDOX!S4BHGCQT4!LIBXMFZ! `5"6$%,'!"='`5"='27# R! 29 R!=?"%=?`Z! "K=Z!:IR!;Z9ZR!:GOFSLADR!'Z-ZR!*OXTAGSBR!2ZR! 0FAGMLADR!5Z+ZR!)OEEBR!*Z+ZR!0O\\IHOR!+ZR! +BTO\A[AR!1ZR!0FGG4LADR!,Z3ZR!2 XAGMAHCR!)ZR!:FD[OXJR!*Z)ZR!5GZ '%#')=C !b"??Kc!'FXGFABFS! F]QGFBBOCD!CW!L4CHCDOD%QGCHFO D!JODABF!LFBBFDPFG!*#+!ADS!QGCHFOD!OD!ASIMH!WCGL!CW! L4CHCDOX!S4BHGCQT4Z! 165%76% R! 260 R!=K_%=KaZ! "KKZ!9ALBTFGFR!,Z)ZR!#F[LADR!.Z.ZR!+M[A\\ADR! ,ZR!+HT[AMR!3Z2Z!ADS!3GCCJR!5Z'Z!b"??`c! 1GADBXGOQHOCDAM!AEDCGLAMOH4!OD!L4CHCDOX!S4B HGCQT4!AWWFXHB!',08!EIH!DCH!DFOPTECGODP! PFDFBZ! /&"6'()#='P6)J'165'['1'P R! 94 R!`K?U%`K??Z! "KUZ!:GOBXTR!*ZR!2ODPMFHCDR!8Z*ZR!,CBFBR!0Z+ZR!)CD\AMF\R!(Z-ZR!&AGADPCR!0ZR!,AGJBR!9Z)Z!ADS! :IDADAPFR!/Z-Z!b=>>"c!.WWFXH!CW!HGOQMFH!GF QFAH!F]QADBOCD!CD!XTGC LAHOD!BHGIXHIGF!ADS! F]QGFBBOCD!CW!',08!ADS!DFOPTECGODP!PFDFBR !2(e_!ADS!',V'R!OD!L4CHCDOX!S4BHGCQT4Z! !"=']%7%#'!%#)< R! 74 R!=a"%=?"Z! "K_Z!.GOJBBCDR!,Z!b=>>Uc!*FAM%HOLF!*1%0&*! WCG!&1)!GFQFAH%XCDHAODODP!PFDFBZ! !%#$"JE'!"=' `5"= R! 277 R!``%aUZ!

PAGE 83

! aK! "KdZ!.GOJBBCDR!,ZR!9FSEFGPR!3ZR!&AGF4R!#Z!ADS!+DBN FSR!1Z!b=>>"c!'FXGFABFS !',08!HGADBXGOQH! MFNFMB!OD!L4CHCDOX!S4BHGCQT4!"!H4QF!((+!LIBXMF!WOEFGBZ! `5"6$%,'`5"?$OE'I%E'B",,*7 R! 286 R!""``%""a=Z! "K`Z!#AGADPR!,Z+ZR!VAGODPR!5Z'ZR!2AECIGODR!-Z +Z!ADS!8CGDFMIJR!*Z)Z!b=>>>c!,4CHCDOX! S4BHGCQT4!b',c!QGCHFOD!JODABF !MFNFMB!OD!XCDPFDOHAM! ADS!ASIMH!',!QAHOFDHBZ! R*&'^'\*,' ]%7%# R! 8 R!_>`%_"=Z! "KaZ!1TCGDHCDR!&Z+ZR!V4LFGR!5Z0ZR!2OLLCDBR!mZR! ,X&MAODR!&Z!ADS!,C]MF4R!*Z1ZR!KGSZ!b"??`c! .]QADBOCD!CW!HTF!L4CHCDOX!S4BHGCQT4!&1)!GF QFAH!GFSIXFB!F]QGFBBOCD!CW!HTF!WMADJODP! ',+90!PFDFZ! ()#']%7%# R! 16 R!U>`%U>?Z! "K?Z!8MFBFGHR!1Z*ZR!$HHFDR!+Z'ZR!3OGSR!1Z'Z!ADS! 1AQBXCHHR!2Z5Z!b"??`c!1GODIXMFCHOSF!GFQFAH! F]QADBOCD!AH!HTF!L4CHCDOX!S4BHGCQT4!MCXI B!GFSIXFB!F]QGFBBOCD!CW!',+90Z! ()#']%7%# R! 16 R!U>=%U>dZ! "U>Z!8[OHFJR!+Z.ZR!1CDFMMAHCR!0Z5ZR!&TFDR!'ZR!)IMMO DPB%9ADSMF4R!5ZR!&TFDPR!;Z2ZR!1[OPPFGR!2ZR! 2XTFFH\R!1Z.ZR!&ABANADHR!1ZZR!2HCMMR!,ZR!#CEGFPAR!,Z+Z '%#')=C !b=>>"c!+IHCLAHFS! XCDBHGIXHOCD!CW!TOPT%SFDBOH4!XCLQAGAHONF! LAQB!EFH[FFD!GAHR!TILADR!ADS!LCIBFZ! ]%7",%' I%E R! 11 R!"?K_%"?UKZ! "U"Z!5ADBFDR!)ZR!)GCFDFDR!0Z5ZR!3AXTDFGR!'ZR!5A QR!0Z9ZR!&CFG[ODJFMR!,ZR!$FGMFLADBR!:ZR!NAD! SFD!3GCFJR!VZR!)CTMBXTR!3ZR!0FHHFR!'ZR!0MCLQR!5Z5Z '%#')=C !b"??dc!+EDCGLAM!L4CHCDOX! S4BHGCQT4!QGCHFOD!JODABF!MFNFMB!QG CSIXF!CDM4!LOMS!L4CQAHT4!OD!LOXFZ! ()#']%7%# R! 13 R!K"d% K=UZ! "U=Z!*FSS4R!2ZR!2LOHTR!'Z3ZR!*OXTR!,Z,ZR!-FWFGCNOXT R!5Z,ZR!*FOMM4R!0ZR!'ANOBR!3Z,ZR!1GADR!8ZR! *A4EIGDR!9ZR!3GCDBCDR!*ZR!&GCBR!'Z '%#')=C !b"??dc!,OXF!MAXJODP!HTF!L4CHCDOX!S4BHGCQT4! QGCHFOD!JODABF!SFNFMCQ!A!MAHF !CDBFH!QGCPGFBBONF!L4CQAHT4Z! ()#']%7%# R! 13 R!K=_%KK_Z! "UKZ!3FGIMR!&Z(ZR!,APIOGFR!&Z1ZR!+GCDCNOH\R!,Z 5ZR!)GFFD[CCSR!5ZR!,OMMFGR!&ZR!)FTGLADDR!5ZR! 9CIBLADR!'ZR!,FDSFMBCTDR!,Z.Z!ADS!*FSS4R!2Z !b"???c!',08!SCBAPF!AMHFGAHOCDB!GFBIMH! OD!AHGOCNFDHGOXIMAG!XCDSIXHOCD!AEDCGLAMOHOF B!OD!A!LCIBF!L4CHCDOX!S4BHGCQT4!LCSFMZ! ^' B=57'278%E# R! 103 R!*"%`Z! "UUZ!3FDSFGBR!+Z+ZR!)GCFDFDR!0Z5ZR!$FGMFLADBR!:Z1 ZR!/FFGJALQR!5Z9Z!ADS!VOFGODPAR!3Z!b"??`c! ,4CHCDOX!S4BHGCQT4!QGCHFOD!JO DABF!OB!ODNCMNFS!OD!HTF! LCSIMAHOCD!CW!HTF!&A=l! TCLFCBHABOB!OD!BJFMFHAM!LIBXMF!XFMMBZ! ^'B=57'278%E# R! 100 R!"UU>%"UU`Z! "U_Z!-FFR!9Z&ZR!0AHFMR!,Z8ZR!,OBHG4R!'Z5ZR! VADPR!>Kc!+EDCGLAM!#A!XTADDFM!PAHODP!OD !LIGODF!XAGSOAX!L4CX4HFB!SFWOXOFDH!OD! L4CHCDOX!S4BHGCQT4!QGCHFOD!JODABFZ! /$OE5"=']%7",56E R! 12 R!"U`%"_`Z! "UdZ!0AMMR!)Z2ZR!5CTDBCDR!8Z5Z!ADS!2LOHTR!)Z-Z!b=>>Kc!+EDCGLAM!XCDHGAXHOMF!AXHONOH4!ADS! XAMXOIL!X4XMODP!OD!XAGSOAX!L4CX4HFB! OBCMAHFS!WGCL!',08!JDCXJCIH!LOXFZ! /$OE5"=' ]%7",56E R! 13 R!"K?%"UdZ!

PAGE 84

! aU! "U`Z!8MFBFGHR!1Z*ZR!&TCR!'Z9ZR!&MAGJR!5Z(ZR!,A4M OFR!5ZR!+SFMLADR!5ZR!2DOSFGR!-ZR!;IFDR!.Z&ZR! 2CGOADCR!0Z!ADS!1AQBXCHHR!2Z5Z!b=>>>c!,O XF!SFWOXOFDH!OD!2O]_!SFNFMCQ!XAHAGAXHBf! OLQMOXAHOCDB!WCG!L4CHCDOX!S4BHGCQT4Z! ()#']%7%# R! 25 R!">_%">?Z! "UaZ!2AGJAGR!0Z2ZR!+QQIJIHHADR!3ZR!9ADR!5ZR!(HCR!;ZR! +OR!&ZR!1BAOR!VZR!&TAOR!;ZR!2HCIHR!5Z1Z!ADS! *FSS4R!2Z!b=>>>c!9FHFGC\4PCIB!MCBB!CW!2O]_!OD !LOXF!OB!BIWWOXOFDH!HC!XAIBF!CXIMAG! XAHAGAXHBZ! ()#']%7%# R! 25 R!"">%""UZ! "U?Z!2AGJAGR!0Z2ZR!0AIMR!2ZR!9ADR!5Z!ADS!*FSS4R!2Z !b=>>Uc!2O]_!OB!GFYIOGF S!WCG!BQFGLAHCPFDOX! XFMM!BIGNONAM!ADS!BQFGLOCPFDFBOBZ! \*,'!"=']%7%# R! 13 R!"U="%"UK"Z! "_>Z!VAJOLCHCR!9ZR!,APIOGFR!&Z1ZR!2TFG[CCSR!,Z &ZR!/AGPABR!,Z,ZR!2AGJAGR!0Z2ZR!9ADR!5ZR! *FSS4R!2Z!ADS!3FGIMR!&Z(Z!b=>>=c!&TAGAXH FGO\AHOCD!CW!XAGSOAX!XCDSIXHOCD!B4BHFL! AEDCGLAMOHOFB!OD!LOXF![OHT!HAGPFH FS!SOBGIQHOCD!CW!2O]_!PFDFZ! ^'27#%&8'B)&J'R=%6#&"?$OE5"= R! 7 R!"=`%"K_Z! "_"Z!:AGJAB%3AGPFHCDR!.ZR!3AGEFHR!5Z0ZR!'ADXFAR!2ZR!VFTGMFR!*ZR!&TFXCIGOR!+Z!ADS!'IMAXR!$Z! b"?aac!(LLAHIGOH4!CW!LIBXMF!WOEFGB!OD!HT F!XCDPFDOHAM!WCGL!CW! L4CHCDOX!S4BHGCQT4f!OHB! XCDBFYIFDXFB!ADS!OHB!CGOPODZ! ^'(%*&"='165 R! 83 R!"U_%"_?Z! "_=Z!2ATPAMR!/ZR!3FGDFBR!2ZR!2ATPAMR!2ZR!-OBXT[F 4R!&Z!ADS!2IEGALADOR!/Z!b"?aKc!2JFMFHAM! LIBXMF!OD!QGFHFGL!ODWADHB![OHT!XCDPFD OHAM!L4CHCDOX!S4BHGCQT4Z!,CGQTCMCPOX!ADS! TOBHCXTFLOXAM!BHIS4Z! ^'(%*&"='165 R! 59 R!U`%__Z! "_KZ!2AGDAHR!9Z3Z!ADS!2OMEFGHR!2ZVZ!b"?`dc!,AHI GAHOCDAM!AGGFBH!CW!WFHAM!LIBXMF!OD!DFCDAHAM! L4CHCDOX!S4BHGCQT4Z!+!QAHTCMCP OX!BHIS4!CW!WCIG!XABFBZ! P&6$'(%*&"= R! 33 R!Udd%U`UZ! "_UZ!3G4BCD%*OXTAGSBCDR!*Z5Z!ADS!&I GGOFR!0Z'Z!b=>>ac!1TF!PFDFHOXB !CW!NFGHFEGAHF!L4CPFDFBOBZ! ()#'I%8']%7%# R! 9 R!dK=%dUdZ! "__Z!9AGQFGR!0Z2Z!b"?a?c! !O"#"756'JOE#&"?$O Z!=DS!FSZ!2AIDSFGBR!-CDSCD!g!0TOMASFMQTOAZ! "_dZ!VADBODJR!'Z)Z!ADS!VOFGODPAR!3Z!b=>>Kc !1GADBPFDOX!LCIBF!LCSFMB!WCG!L4CHCDOX! S4BHGCQT4!H4QF!"!b',"cZ! BO#"D%7%#']%7",%'I%E R! 100 R!=K>%=U=Z! "_`Z!:AGSAFOR!,ZR!-AGJODR!8ZR!3GCCJR!5Z'Z!ADS!9ALB TFGFR!,Z)Z!b=>>"c!(D!NONC!XC%MCXAMOBAHOCD! CW!,3#-!QGCHFOD![OHT!',08! F]QADSFS%GFQFAH!HGADBXGOQHBZ! (*6=%56'P65JE'I%E R! 29 R!=`dd% =``"Z! "_aZ!:AGSAFOR!,ZR!*CPFGBR!,Z1ZR!1TCGQFR!9Z,ZR!-A GJODR!8ZR!9ALBTFGFR!,Z)ZR!9AGQFGR!0Z2Z!ADS! 3GCCJR!5Z'Z!b=>>=c!1TGFF!QGCHFODBR!,3#-R!,3 --!ADS!,3e-R!XC%MCXAMO\F!OD!NONC![OHT! DIXMFAG!WCXO!CW!F]QADSFS%GFQFAH!HG ADBXGOQHB!OD!',"!ADS!',=!XFMMBZ! \*,'!"=']%7%# R! 11 R! a>_%a"UZ! "_?Z!*ADILR!-Z0Z!ADS!&CCQFGR!1Z+Z!b=>>dc! *#+%,FSOAHFS!#FIGCLIBXIMAG!'OBCGSFGBZ! P77*' I%8'(%*&"E65 Z!

PAGE 85

! a_! "d>Z!&TCR!'Z9ZR!1TOFDFBR!&Z0ZR!,ATCDF4R!2Z.ZR!+DAMAIR!.ZR!:OMOQQCNAR!)Z#Z!ADS!1AQBXCHHR!2Z5Z! b=>>_c!+DHOBFDBF!HGADBXGOQ HOCD!ADS!TFHFGCXTGCLAHOD!AH!HTF!',"!&1)!GFQFAHB!AGF! XCDBHGAODFS!E4!&1&:Z! !"='B%== R! 20 R!UaK%Ua?Z! "d"Z!8I4ILXI%,AGHODF\R!#Z,ZR!VADPR!)Z2Z!ADS!&C CQFGR!1Z+Z!b=>>`c!(DXGFABFS!BHFAS4%BHAHF! MFNFMB!CW!&7)30"!OD!L4CHCDOX!S4B HGCQT4!"!AGF!SIF!HC!08&%LFSOAHFS! T4QFGQTCBQTCG4MAHOCDZ! !"='B%== R! 28 R!da%`aZ! "d=Z!1OLXTFDJCR!#Z+ZR!0AHFMR!*ZR!(AJCNAR!0ZR!&AOR! mZ5ZR!>Uc! $NFGF]QGFBBOCD!CW!&7)!HGOQMFH!GFQFAH%EO DSODP!QGCHFODR!&7)30"R!OD!LOXF!ODTOEOHB! L4CPFDFBOBZ! ^'`5"='B$%, R! 279 R!"K"=?%"K"K?Z! "dKZ!-ASSR!+Z#ZR!2HFDEFGPR!,Z)ZR!2[ADBCDR!,Z2Z! ADS!&CCQFGR!1Z+Z!b=>>_c!'4DALOX!EAMADXF! EFH[FFD!AXHONAHOCD!ADS!GFQGFBBOCD!GFPIMAHFB! QGF%L*#+!AMHFGDAHONF! BQMOXODP!SIGODP!TFAGH! SFNFMCQLFDHZ! +%8'+O7 R! 233 R!`aK%`?KZ! "dUZ!8GCMR!5ZR!:OB\FGR!+ZR!,4JC[BJAR!+ZR!2CEX\A JR!8ZR!SF!,F\FGR!,Z!ADS!8G\4\CBOAJR!VZ5Z! b=>>`c!*OECDIXMFABF!SOXFG!XMFANFB! HGOQMFH!GFQFAH!TAOGQODB!ODHC! BTCGHFG!GFQFAHB!HTAH!BOMFDXF! BQFXOWOX!HAGPFHBZ! !"='B%== R! 25 R!_`_%_adZ! "d_Z!1OADR!3ZR!VTOHFR!*Z5ZR!eOAR!1ZR!VFMMFR!2ZR! 1IGDFGR!'Z9ZR!,AHTF[BR!,Z3Z!ADS!1TCGDHCDR! &Z+Z!b=>>>c!.]QADSFS!&7)!GFQFAH!*#+B!WCGL! TAOGQODB!HTAH!AXHONAHF!HTF!SCIEMF%BHGADSFS! *#+%SFQFDSFDH!QGCH FOD!JODABF!08*Z! I(P R! 6 R!`?%a`Z! "ddZ!/ABISFNADR!2ZR!1CDPR!;Z!ADS!2HFOH\R!5Z+Z!b=>>`c !2[OHXTODP!WGCL!GFQGFBBOCD!HC!AXHONAHOCDf! LOXGC*#+B!XAD!IQ%GFPIMAHF!HGADBMAHOCDZ! 165%76% R! 318 R!"?K"%"?KUZ! "d`Z!-OR!-Z&ZR!$JODCR!2Z1ZR!mTACR!9ZR!0CCJCHR!'ZR!0MAXFR!*Z:ZR!7GAJALOR!2ZR!.DCJOSAR!9Z!ADS! 'ATO4AR!*Z!b=>>dc!2LAMM!SB*#+B!ODSIXF!HG ADBXGOQHOCDAM!AXHONAHOCD!OD!TILAD!XFMMBZ! /&"6' ()#='P6)J'165'['1'P R! 103 R!"`KK`%"`KU=Z! "daZ!0MAXFR!*Z:ZR!-OR!-Z&ZR!0CCJCHR!'ZR!#CCDADR! .Z5Z!ADS!'ATO4AR!*Z!b=>>ac!,OXGC*#+%K`K! ODSIXFB!F]QGFBBOCD!CW!PFDFB![OHT!XCLQMFLFDHAG4!QGCLCHFG!BFYIFDXFBZ! /&"6'()#='P6)J' 165'['1'P R! 105 R!"d>a%"d"KZ! "d?Z!-IR!
PAGE 86

! ad! 3($)*+09(&+-!28.1&9! 5ABCD!$6*CIGJF![AB!ECGD!OD!'A4HCDR!$TOCR!OD! "?a>Z!!9F!OB!HTF!CMSFBH!CW!H[C!BCDB!ECGD!HC! -FBMOF!ADS!*FS!$6*CIGJFZ!!5ABCD!AHHFDSFS!$TOC !7DONFGBOH4!WGCL!"???%=>>K![TFGF!TF!BHISOFS! EOCMCP4!ADS!QB4XTCMCP4!ADS!FAGDFS! A!3AXTFMCG!CW!2XOFDXF!OD! LOXGCEOCMCP4Z!!+WHFG!PGASIAHOCDR! 5ABCD!LCNFS!HC!)AODFBNOMMFR!:MCGOSA!ADS!iCODFS !HTF!ODHFGSOBXOQMODAG4!QGCPGAL!OD!EOCLFSOXAM! BXOFDXFB!AH!HTF!7DONFGBOH4!CW!:MCGOSA!&CMMFPF!CW !,FSOXODFZ!!5ABCD!SOS!TO B!PGASIAHF![CGJ!OD!'GZ! ,AIG4!2[ADBCD6B!MAECGAHCG4!CW !HTF!,CMFXIMAG!)FDFHOXB!ADS! ,OXGCEOCMCP4!'FQAGHLFDH!ADS! XCLQMFHFS!TOB!0TZ'Z!SOBBFGHAHOCD! OD!,A4!=>>?Z!!5ABCD!QMADB!HC!LCNF !HC!'AMMABR!1F]ABR!OD!HTF! BQGODP!CW!=>>?!HC!EFPOD!TOB!QCBHSCXHCGAM!BHISOFB!OD! 'GZ!.GOX!$MBCD6B!MAECGAH CG4!AH!HTF!7DONFGBOH4! CW!1F]AB!2CIHT[FBHFGD!,FSOXAM!&FDHFGZ!!!!