FIGURE 2.

Rictor requirement for type II IFN-inducible, mTOR-mediated signaling events. A and B, serum-starved U937 cells transfected with control siRNA or Rictor siRNA were treated with human IFNγ for the indicated times. Cell lysates were prepared, and proteins were resolved by SDS-PAGE and then processed for immunoblotting with anti-Ser(P)-473 AKT (A) and anti-Thr(P)-308 AKT, anti-Rictor, and anti-GAPDH antibodies (B) as indicated. The blots were stripped and probed with an anti-AKT antibody. Bands from three independent experiments were quantified by densitometry. Data are expressed as mean ± S.E. of ratios of phospho-AKT over total AKT for each experimental condition. Statistical analysis was performed using Student's t test as indicated. *, p < 0.05. C, serum-starved U937 cells transfected with control siRNA or Rictor siRNA were treated with human IFNγ for the indicated times. Lysates were analyzed by SDS-PAGE and immunoblotted with an anti-Thr(P)-389 p70S6K antibody. The blot was then stripped and reprobed with an antibody against p70S6K.