Sustained Release of Prostaglandin E2 in Fibroblasts Expressing Ectopically Cyclooxygenase 2 Impairs P2Y-Dependent Ca2+-Mobilization
Effect of thapsigargin on Ca2+-mobilization and migration of MEFs in response to UTP and chemotactic stimuli. WT or COX-2 KI cells were activated, or not, for 24 h with LPS (200 ng/mL) plus cytokines (IFN-, TNF-, and IL-1, 20 ng/mL) and then treated for 5 min with 500 nM thapsigargin and in the absence or presence of 5 μM PGE2. The levels of the indicated phosphoproteins were determined by Western blot (a). The capacity of these cells to migrate in transwell was determined after incubation with 5 μM PGE2 and/or 100 μM UTP. The migration was measured after 24 h of incubation in the absence or presence of different combinations of 10% FBS, PGE2 (5 μM), or UTP (100 nM) in the lower wells (b). Results show a representative blot (a) out of three or the mean + SD of four experiments (b). ; versus the same condition in the absence of treatment in the upper chamber.
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