Figure 4.

mTOR inhibition by rapamycin, AZD8055, and Torin-1 influences several protein in the mTOR pathway. MTT cells were treated with rapamycin (A), AZD8055 (B), or Torin-1 (C) at different concentrations (100 nM, 1000 nM, and 3000 nM for rapamycin; 10 nM, 100 nM, and 1000 nM for AZD8055 and Torin-1). The cells were incubated in the presence of the drugs for 6 hours. Total cell lysates were prepared and analyzed by Western blotting. Control cells were treated with DMSO alone for the same period of time. The levels of phosphorylated forms of proteins (phospho [p]-Akt[Ser473], p-S6[Ser235/236], p-S6[Ser240/244], p-eEF2k, and p-eIF4B) decrease with increasing concentrations of the drugs. Actin was used as a loading control.