Effect of rapamycin on mTORC1/mTORC2 signaling. DHL-6 and Ly7 cells were treated with 50 nM rapamycin for 2, 4, and 8 hours (A) or for 24 hours (B) with the indicated rapamycin concentration. Phosphorylation of Akt, eIF4E, S6, 4EBP1, and mTOR was assessed by Western blot analysis. (C) Malignant cells from DLBCL patients (n = 4) were treated with 50 nM rapamycin for 6 hours before Akt, eIF4E, and S6 phosphorylation was analyzed by Western blot. (D) Levels of PAkt and p-eIF4E were evaluated in peripheral blood samples from 3 patients (n = 3) with aggressive lymphoma treated in the clinic with single-agent temsirolimus.