Description:
FarnesoidXreceptor(FXR,NR1H4)isamemberofthenuclearhormonereceptorsuperfamily.Thesenuclearhormonereceptorsareligand-activatedtranscriptionfactorsthatelicittheiractionsbybindingtohormoneresponseelements(HREs)inthepromotersoftargetgenesandregulatingtranscriptioninresponsetolipophilicligands.Principle:
FarnesoidXreceptor(FXR,NR1H4)isamemberofthenuclearhormonereceptorsuperfamily.Thesenuclearhormonereceptorsareligand-activatedtranscriptionfactorsthatelicittheiractionsbybindingtohormoneresponseelements(HREs)inthepromotersoftargetgenesandregulatingtranscriptioninresponsetolipophilicligands.
Similartoothernuclearreceptors,whenactivated,FXRtranslocatestothecellnucleus,formsadimerwithRXRandbindstohormoneresponseelementsonDNA,whichup-ordown-regulatestheexpressionofcertaingenes,involvedinlipidandglucosehomeostasiswhichplayacrucialroleinpathophysiologyofmanymajordiseasessuchasdiabetes,obesity,atherosclerosisandheartfailure.Inaddition,FXRactivationhasbeenshowntobecriticalintheregulationofinflammatoryresponses.
SignosisnowoffersFXR-Luciferasereporterstablecelllinetotheresearchcommunity.Thishigh-qualitystablecelllineswillfacilitatefurthermolecularstudiesofFXRpathwayitsfunctions.
PrinciplebehindTFluciferasereporter. TFluciferasereporterstablecelllineutilizesartificialpromoterconstructstodriveluciferaseexpression. Thepromoterregioncanconsistsofmultiplerepeatsofacis-elementTFbindingsite,aDNAfragmentfromthepromoterregionofaknownTFdownstreamgene,oraDNAfragmentcontainingputative/knownTFbindingsites. ThereareseveralwaysthataTFcanbeactivated,suchasthroughextracellularstimuliorthroughintracellularsignalingpathways. Onceactivated,theTFtranslocatestothenucleusandofteninteractswithrelevantco-factorstodrivegeneexpression. Onceluciferaseisexpressed,itcangeneratelightinanenzymaticassayandtheamountoflightmeasuredispositivelycorrelatedwiththelevelofTFactivation. |
Data:
AnalysisoftheFXRReporterHepG2StableCellLineinresponsetostimuli.Thecellswereseededona96-wellplateovernightwithDMEMincluding10%FBS.Thecellsthenweretreatedwithorwithout20μMGW4064,10μMGW4064and0μMGW4064respectivelyinDMEMand0.1%FBSfor16hours.Readsaremorethattenfoldhigherthancontrol.
