Figure 1. Activation of γ1 versus γ2 AMPK complexes by 991.

HEK293T cells were co-transfected with cDNA encoding either γ1 or γ2 together with α1 or α2 and β1 or β2. Thirty-six hours post-transfection, cells were serum-starved for 2 h and incubated with DMSO or 991 (1 μM) for 30 min prior to cell lysis. (A) Representative blots showing Thr¹⁷² phosphorylation and β1/β2 expression, determined by western blotting using an automated capillary-based electrophoresis system (Simon, Protein Simple), are shown. (B) Graphs showing quantification of Thr¹⁷² phosphorylation relative to β-subunit expression. (C) Thr¹⁷² phosphorylation and β2 expression were determined in response to varying concentrations of 991. (D) Fold change in Thr¹⁷² phosphorylation: β-subunit expression relative to the corresponding γ1-complex in the absence of 991 is shown. Results shown for (B) and (D) are mean ± SEM for two independent experiments.