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1、RegulationofmalegermcellcyclearrestanddifferentiationbyDND1ismodulatedbygeneticbackgroundDEV057000SupplementaryMaterialFilesinthisDataSupplement:?SuDDlementalTableS1??SuDDlementalFiqureS1一zr岳HavxFig.S1.Neoplasiaformationin129/SvJmutanttestes?E-cadherin(red
2、).129/SvJmutantgermcellsexpressECAD,MVH,NANOG,SOX2andOCT4atE13.5(A-D,arrows).ByE16.5,ECAD-positiveneoplasiasappearinsidetestiscordsandexpressthepluripotencymarkersNANOG,SOX2andOCT4,butnotthegerm-celldifferentiationgeneMVH(E-H)?Atthisstage,therearestillafew
3、isolatedandmorphologicallynormalgermcellsthathavefaintornoMVHexpression(E,inset,arrowheads)butexpressthepluripotentmarkersNANOGandSOX2(F,G,arrowheads).ByE19.5,theseECAD-positivemassesgrowintonascentteratomas.Pluripotentgeneexpressionisstilldetected,restric
4、tedtopocketsofcellswithinthetumor(l?L).Scalebars:50um.?SuDDlementalFigureS2-AIBcDE、f「/F?、y?/G、V/?i?L?■*/EXADHIRIVOc4EGFPNANOGMERGEFig.S2?OriginandgrowthofneoplasiasinDnd1Ter/TerandDnd1Ter/+129/SvJOct4-EGFP-positivetestes.(A-D)E15.5Dnd1Ter/Termutanttestis.E
5、CAD-positive(red)neoplasiasbegintoform,whichareGFP-(green)andNANOG-(blue)positive?Evencellsthatmorphologicallyresemblewild-typegermcellsareNANOG-positiveatthisstage(unlikewild-typecells).(E-H)E19.5Dnd1Ter/^heterozygotewithaneoplasia?Atthisstage,manyisolate
6、dgermcellsareGFP-positive,NANOG-negative,andstilllookmorphologicallynormal(arrowheads).TheECAD-positiveneoplasiahasescapedfromthetestiscordsandgrownmuchlarger.ExpressionofGFPandNANOGarerestrictedtopocketsofcellswithinthegrowingtumor(arrows).Thisphenotypere
7、semblesthehomozygousDnd1Ter/Termutantsonamixedgeneticbackground,wheresomegermcellsretaingermcellidentityatlaterstages(comparewithFig.1D-F).?SuDDlementalFigureS3一E14.5Sox9,Cyp26bl,Fgf9,DhhExpression■Ter+/+■Ter+/-Ter-/-Fig.S3.Themalesomaticprogramisinitiated
8、normallyinmutantgonadsbasedonestablishedmarkers.Comparedwithcontrols,mutantgonadsexpresssimilarlevelsofimportantmalesomaticmarkerssuchasSox9、Cyp26b1,Fgf9andDhh.ControlexpressionofOct4indicatesthatmutantshaves
