Figs1.

Supplemental II – Supplemental Figures containing GS protein expression, GS maximal activity, PDH-Eα phosphorylation (site 1 and 2) and expression and representative western blots. Blood samples were obtained before the clamp (−30 min and 0 min) and every 20 min during the clamp and analyzed for plasma FFA concentration (Fig. S1). Substrate selection was evaluated at a whole body level as RER based on indirect calorimetry (Fig. S2). Muscle biopsies were obtained before (basal state) and under insulin-stimulated conditions (following 120 min insulin infusion and investigated for protein expression of glycogen synthase (GS) (Fig. S3) as well as enzyme activity of GS under saturating and hence maximal conditions (in the presence of 8 mM G6P) (Fig. S4), PDH-Eα site 1 and 2 phosphorylation (Figs. S5 and S6) and PDH-Eα protein expression (Fig. S7). Muscle samples collected during the human glycogen supercompensation regime were investigated for phosphorylation and expression of key metabolic proteins by use of immunoblotting with site-specific antibodies (n = 8–9) and representative blots are given in Fig. S8. Protein expression of mitochondrial proteins in quadriceps muscle from WT and inducible AMPK α double KO mice (imdKO) was investigated by immunoblotting of muscle lysates (n = 6–8) and a representative western blot is provided in Fig. S9. ^∗p ≤ 0.5 and ^∗∗∗p ≤ 0.001 for effect of insulin infusion/time. ^†p ≤ 0.005 and ^†††p ≤ 0.001 for effect of leg. ^¤¤p ≤ 0.01 for significant different from day 1. Main effect is given by horizontal line. Coomassie staining was used to verify even protein transfer.