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Signosis/Stat3 Luciferase Reporter Hela Stable Cell Line/SL-0003-FP/1 Ea
Description:
STAT3ResponsiveLuciferaseReporterHelaStableCellLineisderivedfromhumancervicalcancer,andstablyexpressfireflyluciferasereportergeneunderthecontroloftheSTAT3responseelement. ThiscelllineisanidealcellularmodelformonitoringtheactivationofJAK-STATReceptorSignalingPathwaytriggeredbystimulitreatment,enforcedgeneexpressionandgeneknockdown.Principle:
STAT3isactivatedinresponsetovariouscytokinesandgrowthfactorsincludingIFNγ,IL-6andOSM.TheactivationofSTAT3resultsinformationofSTAT3homodynesandSTAT3/STAT1heterodimersthattranslocatetothecellnucleusandinducetranscriptionofgenesbybindingtotheconsensuselementonthepromoterregionoftargetgenesassociatedwithcellgrowthandapoptosis.SignosishasestablishedStat3luciferasereporterHeLastablecellline,whichcanbeused asareportersystemformonitoringtheactivationofStat3triggeredbystimulitreatment,enforcedgeneexpressionandgeneknockdown.
Thecelllinewasestablishedbytransfection usinga pTA-Stat3-luciferasereportervector,whichcontains4repeatsofStat3bindingsites,aminimalpromoterupstreamofthefireflyluciferasecodingregion, alongwithhygromycinexpressionvectorfollowedbyhygromycinselection. Thehygromycinresistantclonesweresubsequentlyscreenedforoncostatininducedluciferaseactivity.Theclonewiththehighestfoldinduction(10fold)wasselectedandexpandedtoproducethisstablecellline.
PrinciplebehindTFluciferasereporter. TFluciferasereporterstablecelllineutilizesartificialpromoterconstructstodriveluciferaseexpression. Thepromoterregioncanconsistsofmultiplerepeatsofacis-elementTFbindingsite,aDNAfragmentfromthepromoterregionofaknownTFdownstreamgene,oraDNAfragmentcontainingputative/knownTFbindingsites. ThereareseveralwaysthataTFcanbeactivated,suchasthroughextracellularstimuliorthroughintracellularsignalingpathways. Onceactivated,theTFtranslocatestothenucleusandofteninteractswithrelevantco-factorstodrivegeneexpression. Onceluciferaseisexpressed,itcangeneratelightinanenzymaticassayandtheamountoflightmeasuredispositivelycorrelatedwiththelevelofTFactivation. |
Data:
Figure1:AnalysisofHeLa/STAT3-luciferaseactivityofcontrolandtreatmentwithOncostatin. Thecellswereseededona96-wellplateforovernightwithDMEMincluding10%FBS.Thecellsthenweretreatedwithorwithout10ng/mlOncostatinrespectivelyinDMEMand0.1%FBSfor8hours. Stat3luciferasereportercelllineshowedmorethan25-foldincreaseinluciferaseactivityinresponsetoOncostatinMtreatment.