Figure 3.

AKT inhibition decreases endogenous BCA2 protein levels, followed by increased AMPK activation. MDA MB 231 cells were treated with LY294002 in either a dosing study (A; 12-hour time point) or a kinetics experiment (B; 10 µm). The included figures are representatives of experiments done in triplicates, and the included densitometry is the mean ± SEM of phosphorylated proteins (normalized to total) and BCA2 to β-actin, compared to vehicle control (set to 1) for fold change expression. The prepared cell extracts in each experiment were used for Western blots with specific antibodies to pAKT (S473), BCA2, pAMPKα (Thr172), total AMPKα1, and β-actin as loading control. LY, LY294002.