Figure 3. Activities of inflammatory enzymes involved in mediating the osmotic expression of the NFAT5 gene in RPE cells. The level of NFAT5 mRNA was determined with real-time RT–PCR analysis in cells cultured 6 h in iso- (control) and hyperosmotic (+100 mM NaCl) media. The following blocking agents were tested: the cell-permeable reducing agent dithiothreitol (DTT; 300 µM); the reactive oxygen species ROS inhibitor, N-acetyl-L-cysteine (NAC; 1 mM); the inhibitor of mitochondrial permeability transition, cyclosporin A (CsA; 1 µM), the phospholipase A2 (PLA₂) inhibitor, 4-bromophenacyl bromide (Bromo; 300 µM); and the cyclooxygenase (COX) inhibitors acetylsalicylic acid (ASA; 2 mM) and ibuprofen (400 µM). Means ± standard error of the mean (SEM) of 3–5 independent experiments using cell lines from different donors. Significant difference versus unstimulated control: *p<0.05. Significant difference versus NaCl control: ●p<0.05.