Fig. 7.

991 and contraction additively increase AMPK activity and glucose transport in mouse skeletal muscle. EDL muscles from C57BL/6 mice were isolated and incubated in the presence or absence of 991 (5 μM) for 45 min. Muscles were then either electrically stimulated to induce contraction or further incubated (no electrical stimulation) for 15 min. 2-DG transport was measured during the last 10 min of contraction or incubation as described in materials and methods. A: IB analysis was performed on 20 μg muscle lysate protein with the indicated antibodies. Representative blots are shown; n = 8. Shorter (20 s) and longer (2 min) exposure indicate the exposure time with X-ray film. B: 2-DG transport (n = 8). C and D: AMPK complexes were immunoprecipitated from 200 μg muscle lysate protein using either AMPKγ1 (C) or AMPKγ3 (D) antibody. AMPK activity was measured as described in materials and methods. Results are expressed as means ± SE. *Significance of contraction versus the respective control condition (0 or 5 μM 991); #significance of 991 versus the respective vehicle condition with or without 1-Hz contraction. P < 0.05.