Sustained Release of Prostaglandin E2 in Fibroblasts Expressing Ectopically Cyclooxygenase 2 Impairs P2Y-Dependent Ca2+-Mobilization
Characterization of EP1–4 and P2Y2–P2Y4 expression and effect of ionophores on Ca2+-mobilization in MEF cells. The expression levels of the prostaglandin receptors EP1–4 and the levels of P2Y2 and P2Y4 were determined by qPCR (a-b). The response to 1 μM ionomycin (c) and 500 nM thapsigargin (d) on Ca2+-mobilization was determined in MEFs overexpressing COX-2, using the dual excitation 340/380 nm protocol as described in Section 2. MEFs KI were washed with fresh medium to remove PGE2 accumulated and maintained in the absence or presence of 1 μM DFU and 5 μM PGE2. Different extracellular concentrations of calcium were used. Results show the mean + SD of three experiments (a-b) or a representative trace (c-d). versus the same condition in WT cells.
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