Fig. 2.

Impaired insulin (Ins)-induced Akt phosphorylation accompanied with abnormal inducible nitric oxide synthase (iNOS) expression in GK VSMC. Equal amounts of protein from WKY and GK cell lysate sample were resolved by SDS-PAGE and transferred to nitrocellulose membrane. A: Western blot analysis for protein expression. B: intensities of iNOS protein were quantitated by densitometric analysis and normalized to the abundance of total β-actin. Quiescent VSMCs at day 9 were stimulated with insulin (1, 10, and 100 nM) for 10 min (n = 3). C: equal amounts of protein from each cell lysate sample were resolved by SDS-PAGE, transferred to nitrocellulose membrane, and the total and phosphorylated levels of Akt and iNOS expression measured by Western blot analysis. Intensities of phosphorylated protein were quantitated by densitometric analysis and normalized to the abundance of total protein for Akt and to β-actin for iNOS. D: insulin dose-response curve versus Akt phosphorylation were plotted as fold increase. E: insulin dose-response curve versus iNOS expression were plotted. *P < 0.05 WKY vs. GK.