N-terminalFLAG-taggedratPDE7Ahasbeenstablyexpressedinahumanembryonickidney(HEK293)celllineanditsexpressionwasconfirmedbyWesternblotting.TheregulationofintracellularlevelofcAMPbyratPDE7AinratPDE7Astably-expressedHEK293cellswascharacterizedbyacell-basedreporterassayusingpCRE-lucreportervector.pCRE-luccontainsaluciferasegenethatisunderthecontrolofthecAMPresponseelement(CRE).WhencellstransientlytransfectedwithpCRE-lucreporterwereactivatedbyforskolin,thelevelofcAMPwasupregulatedinparentalHEK293cellsinducingtheexpressionofluciferasereporter,whereasratPDE7A-HEK293cellsshowedreductioninthelevelofcAMP,resultinginloweredlevelsofluciferaseexpression.InhibitionofPDE7AactivitybyBRL50481,aPDE7Ainhibitor,restoredthecAMPlevel,resultinginhigherluciferaseactivity.
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2.FanChung,K.(2006).Eur.J.Pharmacol.533(1-3):110-117.
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