Description:
StemcellsareimportantcellscharacterizedbytheirABIlitytoself-renewordifferentiateintomanycelltypes,whichisgovernedbyintracellularsignalingpathwayandtranscriptionregulation.Hence,transcriptionregulationplaysadeterminingroleinconferringcellularidentityandfunction.Thestep-wisematurationofstemcellsintoterminallydifferentiatedcelltypesrequiresthetimelyactivationofacascadeoftranscriptionprogramsgovernedbylineage-specifyingtranscriptionfactors.Multipletranscriptionfactorsandtargetgeneshavebeenwidelyreportedtoassociatewithstemcellself-renewalandpluripotencyincludingEGR1,OCT4,FOXD3,FOXO,Nanog,SOX2,SOX18,ETS,GLI,KLF4,MEF2,Myc,RNUX1,Pax6,TCF/LEFandGATA.However,stemcellsarepreciousandlimited.ApplicableGrid:
ListofApplicableTFs:
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |
| A | EGR1 | Ets | EGR1 | Ets | EGR1 | Ets | EGR1 | Ets | EGR1 | Ets | EGR1 | Ets |
| B | FOXD3 | Gli | FOXD3 | Gli | FOXD3 | Gli | FOXD3 | Gli | FOXD3 | Gli | FOXD3 | Gli |
| C | FOXO1 | KLF4 | FOXO1 | KLF4 | FOXO1 | KLF4 | FOXO1 | KLF4 | FOXO1 | KLF4 | FOXO1 | KLF4 |
| D | GATA | MEF2 | GATA | MEF2 | GATA | MEF2 | GATA | MEF2 | GATA | MEF2 | GATA | MEF2 |
| E | Nanog | Myc | Nanog | Myc | Nanog | Myc | Nanog | Myc | Nanog | Myc | Nanog | Myc |
| F | OCT4 | RNUX1 | OCT4 | RNUX1 | OCT4 | RNUX1 | OCT4 | RNUX1 | OCT4 | RNUX1 | OCT4 | RNUX1 |
| G | SOX2 | PAX6 | SOX2 | PAX6 | SOX2 | PAX6 | SOX2 | PAX6 | SOX2 | PAX6 | SOX2 | PAX6 |
| H | SOX18 | TCF/LEF | SOX18 | TCF/LEF | SOX18 | TCF/LEF | SOX18 | TCF/LEF | SOX18 | TCF/LEF | SOX18 | TCF/LEF |
Principle:
Signosis’stemcellTFactivationprofilingplatearrayisusedformonitoringtheactivationofmultipleTFssimultaneously. Inthetechnology,aseriesofbiotin-labeledprobesaremadebasedontheconsensussequencesofTFDNA-bindingsites. Whentheprobemixincubateswiththecelllysate,individualprobeswillfinditscorrespondingTFandformTF/probecomplexes,whichcanbeeasilyseparatedfromfreeprobesthroughsimplespincolumnpurification. Theboundprobesaredetachedfromthecomplexandanalyzedthroughhybridizationwithaplate;eachwellisspecificallypre-coatedwithcomplementarysequencesoftheprobes.ThecapturedDNAprobeisfurtherdetectedwithstreptavidin-HRP.Luminescenceisreportedasrelativelightunits(RLUs)onamicroplateluminometer.
Data:
StemcellTFactivationprofilingplatearrayanalysisofTFactivitiesinHeLaandStemcellF9.ThewholecelllysateswerepreparedandsubjectedtoTFprofilingassay.
