Fig. 4.

Effect of NOS and Akt inhibitors on H₂O₂-induced NO generation. HCMVEC on coverslips were pretreated with the NOS inhibitor N^ω-nitro-l-arginine methyl ester (l-NAME; 200 μM) or the Akt inhibitor Akt/PKB signaling inhibitor-2 (API-2; 10 μM) for 2 h and then treated with 500 μM H₂O₂ for 10 min. Cells were washed with endothelial basal medium-2 (EBM-2), and coverslips were mounted in the perfusion chamber and perfused with EBM-2 containing 1 μM 4,5-diaminofluorescein diacetate (DAF-2 DA). DAF-2 fluorescence was recorded at 30 min with excitation of 480 nm and emission of 520 nm. Fluorescence intensity was quantified using MetaMorph software.