Description:

NRF2/AREResponsiveLuciferaseReporterNIH3T3StableCellLineisderivedfromMousefibroblast,andstablyexpressfireflyluciferasereportergeneunderthecontroloftheNRF2/AREresponseelement. ThiscelllineisanidealcellularmodelformonitoringtheactivationofAntioxidantResponsePathwaytriggeredbystimulitreatment,enforcedgeneexpressionandgeneknockdown.

Principle:

NRF2playsacrucialroleincellularanti-oxidantdefense,makingitatherapeutictargetforneurodegenerativediseasesandcancer. Undernormalconditions,NRF2localizesinthecytosolandisrapidlydegradedbytheproteasome. Underoxidativestress,NRF2isstABIlizedandtranslocatestothenucleuswhereitbindstoaDNApromoterandinitiatesgeneexpression. Inthenucleus,NRF2formsaheterodimerwithasmallMafproteinandbindstotheAntioxidantResponseElementintheupstreampromoterregionofmanyantioxidativegenes,andinitiatestheirtranscription.

ThisNRF2luciferasereporterstablecelllinehasbeenstablytransfectedwithpTA-ARE-luciferasereportervector,whichcontains4repeatsofantioxidantresponsebindingsites,aminimalpromoterupstreamofthefireflyluciferasecodingregion,alongwitha hygromycinexpressionvector. Followingselection,the hygromycinresistantclonesweresubsequentlyscreenedforTBHQ-inducedluciferaseactivity.Theclonewiththehighestfoldinductionwasselectedandexpandedtoproducethisstablecellline.

 

PrinciplebehindTFluciferasereporter. TFluciferasereporterstablecelllineutilizesartificialpromoterconstructstodriveluciferaseexpression. Thepromoterregioncanconsistsofmultiplerepeatsofacis-elementTFbindingsite,aDNAfragmentfromthepromoterregionofaknownTFdownstreamgene,oraDNAfragmentcontainingputative/knownTFbindingsites. ThereareseveralwaysthataTFcanbeactivated,suchasthroughextracellularstimuliorthroughintracellularsignalingpathways. Onceactivated,theTFtranslocatestothenucleusandofteninteractswithrelevantco-factorstodrivegeneexpression. Onceluciferaseisexpressed,itcangeneratelightinanenzymaticassayandtheamountoflightmeasuredispositivelycorrelatedwiththelevelofTFactivation.

 

Data:

AnalysisoftheNRF2PathwayReporterNIH3T3StableCellLineinresponsetostimuli. Thecellswereseededona96-wellplatefor8hoursorovernightwithDMEMincluding10%FBS.Thecellsthenweretreatedwithorwithout10μMTBHQor10μM4HNE respectivelyinDMEMand0.1%FBSfor16hours.

  

Signosis基因表达  是利用来自基因的信息指导功能性基因产物合成的过程。这些产品通常是蛋白质。基因表达分析对于理解基因及其蛋白产物的功能，以及对影响基本生物学过程的复杂调控网络的清晰了解至关重要。Signosis  提供了许多用于基因表达研究的测定产品，包括用于监控由TF上调或下调的基因表达的cDNA板阵列，以及用于分析基因表达的单细胞PCR试剂盒，可直接分析细胞无需RNA分离即可进行裂解。 SARS-CoV-2实时PCR检测试剂盒单细胞基因表达分析印迹分析RNA的cDNA板阵列细胞裂解液的cDNA板阵列