Figure 5

Effect of suppression of hepatic GRK2 on insulin signaling in ob/ob mice. (A) Representative western blot analysis of Akt phosphorylation (Ser473 and Thr308), total Akt, eNOS phosphorylation (Ser1177), total eNOS, and β-actin in lean and ob/ob aortas from mice unstimulated or stimulated with insulin (10⁻⁶ mol/L; 20 min). Basically, Insulin-stimulated and non-stimulated lanes were run on the same gel but were noncontiguous, and partly, were run on a separated gel but under the same conditions. (B–F) Quantification proteins are expressed as ratio of Akt to β-actin (B), ratio of eNOS to β-actin (C), ratio of Akt phosphorylation (at Ser473) to total Akt (p-Akt [Ser473]) (D), ratio of Akt phosphorylation (at Thr308) to total Akt (p-Akt[Thr308]) (E), and eNOS phosphorylation (at Ser1177) to total eNOS (p-eNOS [Ser1177]) (F). Phosphorylated proteins are normalized to total proteins. Total proteins are normalized to β-actin. Values are mean ± SE; n = 5–6. ^* p < 0.05, ^** p < 0.01, ^*** p < 0.001 vs. non-stimulated; ^† p < 0.05, ^†† p < 0.01 vs. control siRNA-transfected ob/ob mice (ob/ob-cont si) by one-way analysis of variance.