Figure 2. BEZ235 treatment results in decreased global protein translation. (A) SQ20B or U251MG cells were treated with BEZ235 for 24 h. 30 minutes before the end of treatment, cells were labeled with [³⁵S]methionine (50 µCi/ml). At completion of assay, cells were lysed and assessed for total protein content and radioactivity. Results are represented as the counts per minute (cpm) normalized to total protein concentration in the drug-treated cells compared to the untreated control cells. (B) The effect of PI3K/Akt/mTOR inhibition on cap dependent translation. The figure shows the interactions between eIF4E, 4E-BP1 and eIF4G. (C) SQ20B or U251MG cells were treated with drugs as indicated for 3 h. Whole cell extracts were prepared and eIF4E and its associated proteins purified by 7-methyl-GTP-Sepharose affinity chromatography. (D) SQ20B or U251MG cells were treated with 4EGI1 or BEZ235 for 24 h. After the drug treatment cells were exposed to hypoxia for 3 h. Thereafter, protein was harvested. For panels C and D, protein samples were analyzed by SDS-PAGE and immunoblotting for proteins using antibodies as shown.