Effect of R-568 on Akt activation and NO production in vSMCs from WKY and SHR rats.

(A) vSMCs stimulated with 1μM of R568 (R) at different times (30 seconds, 1 minute, 2 minutes) in the presence or absence of Calhex 231 (C, 1 μM) were subjected to immunoblotting for total and phosphorylated forms of Akt (phospho-Akt^Ser473). Total β-actin levels were used as loading control. A representative set of immunoblots is shown for experiments that were repeated independently 3 times. Each bar represents the mean ± SD of densitometric analysis for phosphorylated proteins normalized to their respective total forms. *p < 0.02 vs respective basal (CTRL), **p < 0.05 R+C vs respective R. (B) NO production determined by conversion of L-[3H]-arginine into L-[3H]-citrulline in vSMCs stimulated with 1μM of R568 in the presence or absence of Calhex 231 (1 μM), data are expressed as pmol/NO/min/mgprottot (picomoles /Nitric Oxide/ minutes/ milligrams protein total). Each bar represents the mean ± SD of 3 independent experiments. *p < 0.02 R568 vs respective basal, **p < 0.01 R568+Calhex vs respective R568.