课件酸谈实验室40读文献学

酸谈辅修课40读文献学实验主讲 猫大信息科 读文献学实验谈谈信号转导（1）……信号转导（2）……磷酸化蛋白信号转导（3）……

转录调控（1）……转录调控转录调控转录调控转录调控（5）……Co- 酸TTF-1regulatesthetotalintracellularcholesterolTTF-1positivelyregulatestheexpressionofthecholesteroleffluxfactorABCA1isadirecttranscriptionaltargetofTTF-TTF-1increasescholesterolUpregulationofTTF-1expressionenhancescellularsensitivitytoFigure1.IntracellularcholesterolcontentisinverselycorrelatedwithTTF-1expressionFigure2.TTF-1regulatesABCA1RNAFigure3.TTF-1regulatesABCA1proteinFigure4.ABCA1promoterreportersaretransactivatedbyTTF-LuciferaseLuciferaseaAdiagramshowstherelativepositionsofLuciferaseLuciferaseResponsesoftheFLreporter(b)andtheindividualfragmentsofABCA1promoter(c)toTTF-1ortheHDDmutantwereinvestigated.dReporterassaysoftheP1fragmentoftheABCA1promotercontainingamutatedTBEasindicated.eAschematicillustratestherelativepositionsofQPCRprobestoputativeTBEsforChIP–QPCRexperiments.fAntibody-pulleddownchromatinswereyzedbyRbrabbit.RLUrelativelightunit,TSStranscriptionstartsite.Dataaremean±SEMofthreeindependentexperiments(*p<0.05;***p<0.001;t-test)Figure5.CholesteroleffluxesvarywithTTF-1/Ttf-1expressionFigure6.TTF-1enhancescellularsensitivityto 酸TTF-1regulatesthetotalintracellularcholesterolTTF-1positivelyregulatestheexpressionofthecholesteroleffluxfactorABCA1isadirecttranscriptionaltargetofTTF-TTF-1increasescholesterolUpregulationofTTF-1expressionenhancescellularsensitivityto 酸Day1Day2Day4裂解细胞，数据测试 酸 酸ExpressionofhumanFCGRTmRNAinhumancellEMSAandantibody ysisoftheputativetranscriptionfactorbindingsitesinthehFCGRTEffectofspecificmutationsinthehFCGRTpromoteronitsFigure2：ElectrophoreticmobilityshiftassayswithprobeSp1-1+2(A)andprobeSp1-3Figure3：IdentificationoftranscriptionfactorsbindingtotheSp1sequenceswithinthe-660/-233fragmentofthehFCGRTpromoter.Figure4：CharacterizationoftheputativeAP-1bindingsiteinthehFCGRTpromoterbyEMSA(A)andsupershift ysis(B).Figure5.CharacterizationoftwopotentialCF1/YY1regulatorymotifsinthe-660/-233fragmentofthehFCGRTpromoterbyEMSA(A,B),andsupershift ysis(A1,B1).Figure6.CharacterizationoftheputativeC/EBPβbindingsiteatposition-497withinthe-660/-233fragmentofthehFCGRTpromoter.Figure7.CharacterizationoftheputativeC/EBPβbindingsiteatposition-233withinthehFCGRTpromoterbyEMSA(A)andsupershift ysis(B).Figure8.RoleofSp1bindingsitesinthepromoteractivityofthehumanFCGRTFigure9.EffectoftheAP-1bindingmotifonthetranscriptionalactivityofthehFCGRTFigure10. ysisoftheCF1bindingsiteswithinthehFCGRTFigure11.EffectofmutationintheC/EBPβbindingsiteonthehFCGRTpromoteractivityinLu106,Caco-2,andHSkMECcelllines.Figure12.EffectofthemutationofC/EBPβmotifsonthehFCGRTpromoteractivityinTHP-1 酸ExpressionofhumanFCGRTmRNAinhumancellEMSAandantibodysupershift ysisoftheputativetranscriptionfactorbindingsitesinthehFCGRTEffectofspecificmutationsinthehFCGRTpromoteronits 酸 酸谈 酸 酸(D)BisulphitesequencingcomparingpromoterDNAmethylationofDNAmethylationdependentgenesinwild-typeandPrdm14-nullEScellsculturedin2iconditions.E Mic60in ctswithTFAMandTFB2M.(A)ImmunostainingofHeLacellswithantibodiesagainsttheantibodiesspecificwithMic60andTFAM.Secondaryred)andFITC(forantibodyagainstTFAM,green)were(blue).Scalebarsindicate10mm.(B)Coimmunoprecipitation(IP)ofMic60andTFAMincelllysatesusingantibodiesspecificforthec-MyctagortheHAtag.CelllysateswereobtainedfromHEK293TcellstransfectedwithMic60-MycandTFAM-HA.(C)Co-IPofendogenousMic60andTFAMinHEK293Tcelllysates.tailofTFAMwasindispensableforthein 5%)withMic60.Thefunctional sofTFAMarerepresentedonthetoprowofpanelD.TFAMCterminal(ct)andTFAMwithaC-terminaltaildeletion(TFAM-cd),whichwasusedfortheinvitrobindingassays,arebinantTFAM-GST,TFAM-cd-GSTorTFAM-ct-GSTproteinsthatwerepreparedbyGSTpurification.Theboundcomplexeswere yzedbyimmunoblotting.