Fig. 2.

Effects of insulin on [³H]2-deoxyglucose uptake (A), and effects of insulin and/or PIP₃ on aPKC activity (B) and phosphorylation of the activation loop site in aPKC (C) in vastus lateralis muscles of high-fat–fed (high fat) and chow-fed (low fat) mice. Mice were injected IP as described in Experimental Procedures (A). aPKCs were immunoprecipitated from muscles of mice treated IM with saline vehicle or insulin (1U/kg) in saline for 15 min, and then assayed in absence or presence of 10 μM PIP₃ (B and C). It was necessary to first immunoprecipitate (IP) aPKC (because other 70–80 kDa PKCs have similar activation loop phosphorylation sites) before Western analyses for phospho-thr-411/410-PKC-λ/ζ; shown here is a blot representative of four determinations (C). Values in panels A and B are mean ± SE of (N) determinations. P values were determined by ANOVA.