Effect of VEGF on COX-1 and COX-2 mRNA and protein expression in HIMEC. Detection of COX-1 and COX-2 mRNA in HIMEC by semi-quantitative reverse transcriptase-PCR using specific primers. HIMEC constitutively express mRNA for COX-1 but not COX-2. Stimulation (3 h) of HIMEC with VEGF (50 ng/ml) led to marked up-regulation of COX-2 gene expression, whereas COX-1 was unaffected. CsA, NS398 and PD098059 pre-treatment of HIMEC inhibited COX-2 mRNA expression following VEGF. β-actin, was used as loading control. Representative figure for HIMEC isolates from five different patients.