FIGURE 2.

Activation of PI3K is sufficient to restore Akt (Ser-473) phosphorylation in cells lacking mTORC2. Sin1^−/− MEFs were (a) treated with rapamycin (100 nm) for the indicated times, (b) treated with DMSO or an active site inhibitor of p70S6K, PF-4708671 (5 μm) for 24 h, (c) transfected with Myr-p110α for ∼30 h, or (d) treated with the indicated doses of Torin1 for 24 h prior to harvest. Immunoblotting was used to determine total and phosphorylated protein levels, and the relative phosphorylation of S6 and Akt (Ser-473), normalized to actin, was determined by densitometry and fit to a dose response curve (kaleidagraph). Data represent the mean ± S.E. of three independent experiments.