Pim and PI3K/AKT/mTOR pathways control phosphorylation of eIF4B S406 and S422, respectively. (A) Cell lines were treated with BEZ235 (0.5 μM), PP242 (1 μM), or AZD8055 (1 μM) for 3 h. Cell lysates were analyzed by immunoblot assays using indicated antibodies. (B) HeLa cells were treated with MK2206 (1 μM), rapamycin (Rapa; 100 nM), PP242 (1 μM), BEZ235 (0.5 μM), U0126 (U0; 15 μM), BI-D1870 (BI; 10 μM), GNE-652 (1 μM), AZD1208 (3 μM), and the indicated inhibitor combinations for 3 h. Cell lysates were analyzed by immunoblot assays using indicated antibodies. (C) HeLa cells were serum starved for 24 h, and then cells were treated with inhibitors as in panel B for 3 h before insulin (1 μg/ml) was added for an additional 30 min. Cell lysates were analyzed by immunoblot assays using the indicated antibodies. (D) PC3-LN4 cells were treated with GSK690693 and GNE-652 as indicated for 24 h. Cell lysates were analyzed by immunoblot assays using the indicated antibodies.