Fig. 3.
The synergistic effect of A769662 and AICAR cotreatment is ablated in AMPKβ1-deficient hepatocytes. Primary hepatocytes were isolated from AMPKβ1−/− mice and wild-type (WT) littermates (12–15 wk old) and cultured overnight. Cells were serum starved for 2–3 h and then treated with vehicle, AICAR, and/or A769662 for 45 min before cell lysis. A: vehicle-treated hepatocyte lysates were analyzed by immunoblotting using 30 μg of protein and the indicated antibodies. B: hepatocyte lysates from AMPKβ1−/− mice were analyzed by immunoblotting using 30 μg of protein extracts and the indicated antibodies. C: AMPKα1 complexes were immunoprecipitated from 30 μg of extract with 1 μg of anti-AMPKα1 and protein G Sepharose. Immune complexes were assayed for AMPK activity (in duplicate). Results are expressed as mean ± SE pmol Pi incorporated·min−1·mg−1; n = 3.