Fig 1.

Effect of incubation with gACRP30 on AMPK and related parameters. EDL muscles from 60-g rats were incubated for 30 min in the presence (+) or absence (−) of gACRP30 (2.5 μg/ml) and then frozen in liquid N₂ until analyzed. (A) AMPK activity in immunoprecipitates obtained with antibody to the α2 isoform. (B) Western blots of AMPK phosphorylated at Thr-172 (active enzyme; p-AMPK), total AMPK abundance (anti-α1 and anti-α2 AMPK antibodies; AMPK), and ACC phosphorylated at Ser-79 (inhibited enzyme; p-ACC). (C) Concentration of malonyl CoA. Blots are representative of muscles of three to five animals. Measurements of AMPK activity and malonyl CoA concentration are means ± SE (n = 5). (D) Effect of gACRP30 on glucose transport in rat EDL. Muscles were incubated with (+) or without (−) gACRP30 (2.5 μg/ml) for 30 min. Glucose transport was then assessed in the absence of insulin based on the uptake of [³H]-2-deoxyglucose. Measurements of glucose transport are means ± SE (n = 6). Additional details are described in Materials and Methods. *, P < 0.05 vs. gACRP30 (−) group.