>
产品中心 >
Stable_cell_line >
Signosis/HIF Luciferase Reporter NIH3T3 Stable Cell Line/SL-0005-FP/1 Ea
Description:
HIFResponsiveLuciferaseReporterNIH/3T3StableCellLineisderivedfrommousefibroblast,andstablyexpressfireflyluciferasereportergeneunderthecontroloftheHIFresponseelement. ThiscelllineisanidealcellularmodelformonitoringtheactivationofHypoxiaReceptorSignalingPathwaytriggeredbystimulitreatment,enforcedgeneexpressionandgeneknockdown.Principle:
Hypoxia-InducIBLeFactor(HIF)-1isadimericproteincomplexthatplaysacentralroleintheresponsetolowoxygenconcentrations,orhypoxia,and isacrucialphysiologicalregulatorofhomeostasis,vascularization,andanaerobicmetabolism.HIFisatranscriptionfactorregulatinggeneexpressionbybindingtotheirDNArecognitionsiteonthetargetgenes.FurThermore,HIF-1hasbeenwidelystudiedbecauseofitsperceivedtherapeuticpotential. HIF-1allowssurvivalandproliferationofcancerouscellsduetoitsangiogenicproperties,andtheinhibitionpotentiallycouldpreventthespreadofcancer.WithagrowingunderstandingoftheHIF-1pathway,ithasbecomeanattractivegoaltoanalyzetheinhibitionandstimulationofHIFtranscriptionalactivityviasmallmolecules.
Thecelllinewasestablishedbytransfection usinga pTA-HIF-luciferasereportervector,whichcontains4repeatsofHIFbindingsites,aminimalpromoterupstreamofthefireflyluciferasecodingregion, alongwithhygromycinexpressionvectorfollowedbyhygromycinselection. ThehygromycinresistantclonesweresubsequentlyscreenedforCoCl2-inducedluciferaseactivity.
PrinciplebehindTFluciferasereporter. TFluciferasereporterstablecelllineutilizesartificialpromoterconstructstodriveluciferaseexpression. Thepromoterregioncanconsistsofmultiplerepeatsofacis-elementTFbindingsite,aDNAfragmentfromthepromoterregionofaknownTFdownstreamgene,oraDNAfragmentcontainingputative/knownTFbindingsites. ThereareseveralwaysthataTFcanbeactivated,suchasthroughextracellularstimuliorthroughintracellularsignalingpathways. Onceactivated,theTFtranslocatestothenucleusandofteninteractswithrelevantco-factorstodrivegeneexpression. Onceluciferaseisexpressed,itcangeneratelightinanenzymaticassayandtheamountoflightmeasuredispositivelycorrelatedwiththelevelofTFactivation. |
Data:
HIFNIH3T3StableCellLineAnalysis. HIFNIH3T3stablecellswereseededina96-wellcultureplatewiththeindicatedmedium. Thecellsweretreatedwith100uMCoCl2for8hours.
