NFATc3-dependent COX2 expression and activity in endothelial cells. (A) shRNA-transformed HUVECs were treated with VEGF (50 ng/mL) for 5 hours and the expression of COX2 mRNA analyzed by qPCR. *P < .05 compared with control (KDSc) (n = 3). (B) Representative Western blot of COX2 protein expression in VEGF-stimulated shRNA-transformed HUVECs. Tubulin (TUB) expression was analyzed as a loading control. (C) EIA detection of prostacyclin release, an index of COX2 activity, in VEGF-stimulated shRNA-transformed HUVECs. Treatment of KDSc-expressing cells with the COX2-specific inhibitor etoricoxib (10μM) was used as a positive control of inhibition. *P < .05 and **P < .01 compared with control (KDSc) (n = 3).