Nutlin 3

Cells were treated with 5 nM ActDL (Sigma-Aldrich, A1410) or 2 μM BMH-21 (Sigma-Aldrich, SML1183) for 24 hours. CHX (Sigma-Aldrich, C4859) was used at a working concentration of 100 μg/ml for 30 min or the indicated time points. For DOX induction (Sigma-Aldrich, D9891), cells were treated with DOX (1 μg/ml; Sigma-Aldrich, D9881) for 48 to 72 hours, unless otherwise indicated. Pan-caspase inhibitor Z-VAD-FMK (R&D Systems, FMK001) and caspase 2 inhibitor Z-VDVAD-FMK (R&D Systems, FMK003) were used at a final concentration of 20 μM for 1 hour or the indicated time points. MG132 (Sigma-Aldrich, M7449), Nutlin-3 (Selleckchem, S1061), and MLN4924 (Selleckchem, S7109) were added to cells in a final concentration of 10, 5, and 1 μM, respectively, for 24 hours. Camptothecin (Sigma-Aldrich, C9911) was used at a working concentration of 5 μM and 5-fluorouracil (Sigma-Aldrich, F6627) at 10 μg/ml. Neocarzinostatin was used at 0.5 μg/ml for 24 hours, LMB was used at 20 ng/ml for 6 hours, and pladB was used at 1 μM for 6 hours. For chemical inhibition of eIF4A3, 52a, or 52b, 53a from (31 (link)) was used at 5 μM for 24 hours. For cell synchronization at G₂, cells were treated with nocodazole (Sigma-Aldrich, M1404) at 400 nM for 16 hours.