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. 2013 Oct 1;18(4):556–566. doi: 10.1016/j.cmet.2013.08.019

Figure 6.

Figure 6

Evidence that ACC Phosphorylation in G361 Cells Is Mediated by AMPK, and Effects of A23187 and Berberine in AMPK Knockout MEFs Expressing α1-T172D Mutant

(A) FLAG-tagged inactive (D157A) mutant of AMPK-α2 was stably expressed by homologous recombination in G361 cells carrying an Flp recombinase target site (see Experimental Procedures) to generate dominant-negative (DN) cells. The graph shows AMPK activity (mean ± SEM, n = 4) measured in immunoprecipitates from control (WT) and DN cells with and without treatment with 10 μM A23187, while the pictures below show results of western blotting to determine expression and phosphorylation of various proteins.

(B) As in (A), except the cells were treated with increasing concentrations of A769662 (30, 100, 300, 500, 1000 μM) or 10 μM A23187.

(C) Effects of A23187 and berberine on AMPK activity and ACC phosphorylation in AMPK KO MEFs that were either untransfected or had been transfected with DNAs encoding myc-tagged AMPK-α1 (wild-type or T172D mutant), AMPK-β2, and AMPK-γ1. Data are mean ± SEM (n = 4). Significant differences from control without A23187 or berberine are shown.