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Description:
NFkBisaubiquitoustranscriptionfactorthatplaysakeyroleincellularresponsestonumerousstimulisuchasstress,cytokines,ultravioletirrADIation,andbacterialorviralantigens.IncorrectregulationofNFkBhasbeenlinkedtocancer,inflammatoryandautoimmunediseases,andsepticshock.WhenNFkBisactivated,itdissociatesfromitsinhibitorIkBandmovesfromthecytoplasmtothenucleus. Onceinthenucleus,itbindstotargetDNAelementsandpositivelyregulatesthetranscriptionofgenesinvolvedinimmuneandinflammatoryresponses,cellgrowthcontrol,andapoptosis.ApplicableGrid:
ListofApplicableGenes
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |
| A | BCL2 | IL6 | Myc | BCL2 | IL6 | Myc | BCL2 | IL6 | Myc | BCL2 | IL6 | Myc |
| B | 18SrRNA | IL9 | NOS2 | 18SrRNA | IL9 | NOS2 | 18SrRNA | IL9 | NOS2 | 18SrRNA | IL9 | NOS2 |
| C | CCND1 | GAPDH | p53 | CCND1 | GAPDH | p53 | CCND1 | GAPDH | p53 | CCND1 | GAPDH | p53 |
| D | Cox2 | IFNg | TNFA | Cox2 | IFNg | TNFA | Cox2 | IFNg | TNFA | Cox2 | IFNg | TNFA |
| E | FASL | IRF1 | TNFR | FASL | IRF1 | TNFR | FASL | IRF1 | TNFR | FASL | IRF1 | TNFR |
| F | INFB1 | MDR1(Abcb1b) | VCAM1 | INFB1 | MDR1(Abcb1b) | VCAM1 | INFB1 | MDR1(Abcb1b) | VCAM1 | INFB1 | MDR1(Abcb1b) | VCAM1 |
| G | IL1A | MMP1 | VEGFC | IL1A | MMP1 | VEGFC | IL1A | MMP1 | VEGFC | IL1A | MMP1 | VEGFC |
| H | IL2 | Myb | Blank | IL2 | Myb | Blank | IL2 | Myb | Blank | IL2 | Myb | Blank |
Principle:
Signosis’proprietaryCDNAplatearrayisaplate-basedhybridizationprofilinganalysisformonitoringtheexpressionofdozensofgenesthroughreversetranscriptionofmRNAintocDNA.Likearrayanalyses,totalRNAisfirstreversetranscribedintocDNAinthepresenceofbiotin-dUTPintheassay.Targetedgenesarethenspecificallycapturedontoindividualwellsonaplate,insteadofmembranes,throughapre-coatedgene-specificoligonucleotide.ThecapturedcDNAsarefurtherdetectedwithstreptavidin-HRP.Luminescenceisreportedasrelativelightunits(RLUs)onamicroplateluminometer.Theexpressionlevelofgenesisdirectlyproportionaltotheluminescentintensity.
Data:
MouseNFkB-regulatedcDNAplatearrayanalysis. NIH3T3cellsweretreatedwith(redbars)orwithout(lightblue)10ng/mlTNFfor30min. Cellswerecollectedforprofiling23geneswiththecDNAplatearrayassay.
