miRNA sponges design supplementary info

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1、MicroRNAsponges:competitiveinhibitorsofsmallRNAsinmammaliancellsMargaretSEbert,JoelRNeilson&PhillipASharpSupplementaryfiguresandtext:SupplementaryFigure1.EffectofspongesonamiR-20targetandanuntargetedcontrol.SupplementaryFigure2.Comparisonofspongestoantisenseoli

2、gos.SupplementaryFigure3.InhibitionofmicroRNAbyastablyexpressedsponge.SupplementaryFigure4.ValidationofnewmicroRNAtargets.SupplementaryTable1.Sequencesofspongesandreporters.SupplementaryTable2.ComparisonofmicroRNAspongestoantisenseoligos.SupplementaryMethodsSup

3、plementaryFigure1120miR-20target100untargetedcontrol80604020Relativeluciferaseunits0C-CXC-20bC-20pfU-CXU-20bU-20pfSpongeSupplementaryFigure1:EffectofspongesonamiR-20targetandanuntargetedcontrol.Wecotransfected293TcellswithaRenillaluciferasevectorcontainingseven

4、miR-20sitesoranotherwiseidenticalvectorcontainingsevenCXCR4controlsitesandthespongeplasmidsindicated.BarsrepresentRenillaluciferaseuntsrelativetoFireflyluciferaseunits.Errorbarsrepresentstandarddeviationamongtriplicatesamples.Resultsarerepresentativeofamimimumo

5、fthreeindependentexperiments.SupplementaryFigure2108642Relativetargetexpression0MockLNA162'-O-me16Sponge16LNAcontrolLNA+sponge2'-O-mecontrolSpongecontrol2'-O-me+spongeSupplementaryFigure2:Comparisonofspongestoantisenseoligos.Wetransfected293Tcellswith20nMantise

6、nseoligo(2'-O-methylorLNA)orwiththeCMVbulgedspongeagainstmiR-16.Negativecontrols:mock(nooligosorsponges),2'-O-methylagainstmiR-30,LNAagainstmiR-122,CXCR4sponge.ThetargetreportercontainsninemiR-16sitesandisderepressedslightlymorestronglybytheLNAandspongethanbyth

7、e2'-O-methyloligo.Errorbarsdenotestandarddeviationintriplicatesamples.Resultsarerepresentativeofaminimumofthreeindependentexperiments.SupplementaryFigure38642RelativemiR-16targetexpression0linelineParental293TCXCR4spongeMiR-16spongeTransienttransfectionSuppleme

8、ntaryFigure3:InhibitionofmicroRNAbyastablyexpressedsponge.Westablytransfected293TcellswiththemiR-16spongeorCXCR4sponge(control)plasmid,sortedforhighGFPexpression,andtestedby