HumanPDE7Bhasbeenstablyexpressedinahumanembryonickidney(HEK293)cellline.PDE7BexpressionwasconfirmedbyWesternblotting.ThefunctionofexpressedPDE7BwasvalidatedbycAMPdetectionassay.TheregulationofintracellularlevelofcAMPbyPDE7BinPDE7Bstably-expressedHEK293cellswascharacterizedbyacell-basedreporterassayusingpCRE-lucreportervector.pCRE-luccontainstheluciferasegenethatisunderthecontrolofcAMPresponseelement(CRE).ElevationofintracellularcAMPactivatescAMPresponseelementbindingprotein(CREB)tobindCREandinducestheexpressionofluciferase.WhencellstransientlytransfectedwithpCRE-lucreporterwereactivatedbyforskolin,thelevelofcAMPwasupregulatedinparentalHEK293cellsinducingtheexpressionoftheluciferasereporterwhereashPDE7B-HEK293cellsshowedreductioninthelevelofcAMPthatresultedinloweredexpressionofluciferase.
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2.FanChung,K.(2006)Phosphodiesteraseinhibitorsinairwaysdisease.Eur.J.Pharmacol.533(1-3):110-117
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