Electrophoretic mobility shift assay of NFATp, NFκB and AP-1 in VEGF and TNF-α/LPS stimulated HIMEC. Nuclear extracts from VEGF (50 ng/ml) or TNF-α/LPS stimulated HIMEC with or without CsA (0.1 μM) were subjected to EMSA using probes to left panel; the NFAT site of human COX-2 promoter (nucleotides -117 to -91). Middle panel; the NFκB site of the human COX-2 promoter (nucleotides -135 to -123). Right panel; the NFAT-AP1 site of the human COX-2 promoter probe (nucleotides -82 to -58). CsA inhibited NFAT DNA binding following VEGF activation in HIMEC (A). NFAT was not activated by TNF-α/LPS. VEGF did not activate NFκB in HIMEC, which was activated following TNF-α/LPS (B). VEGF activated AP-1, which was not inhibited by CsA (C). These data demonstrate the selective inhibitory effect of CsA on NFAT acativation following VEGF stimulation in HIMEC.