Sustained Release of Prostaglandin E₂ in Fibroblasts Expressing Ectopically Cyclooxygenase 2 Impairs P2Y-Dependent Ca²⁺-Mobilization

Figure 2

Characterization of targeting of PKC, PKD, and PKA on the effect of PGE₂ on the UTP-dependent Ca²⁺-mobilization. WT or KI MEFs were washed with fresh medium and maintained in culture for 1 h to remove PGE₂ accumulated and then treated for 10 min with the indicated effectors, except for DFU that was added immediately after washing (1 μM DFU, an inhibitor of COX-2; 5 μM PGE₂; 100 nM staurosporine; 100 nM Gö6976; 5 μM Gö6850; 10 nM Gö6983, a selective inhibitor of classic PKCs; 200 nM PDBU; 200 nM αPDD; 200 nM CID755376, a selective inhibitor of PKD; 5 μM dibutyryl cAMP) and the percentage of cells showing Ca²⁺-mobilization in response to UTP (100 μM) was determined using the nonratiometric Fluo-4 assay. Results show the mean + SD of three experiments for Ca²⁺-mobilization. , versus the same condition in the WT cells.