Effect of EtOH and INK128 on mTORC1/2 activity in DLBCL cells. SUDHL-2 and SUDHL-4 cells were treated with either EtOH for 24 h, INK128 for 3 h at the concentrations as indicated, or 20 nM rapamycin (Rap.) for 3 h. (A) Cell lysates were analyzed by western blotting for phosphorylation states and total levels of indicated proteins. Quantification of the signals is expressed as the percentage of the signal intensity relative to the control group in each cell line. Graphs present the mean and standard deviation from two to five independent experiments. *p ≤ 0.05. (B) The levels of phosphorylated AKT at Ser473 and Thr308, and total AKT levels were assessed. (C) IP assays, using either IgG or anti-mTOR antibodies, were performed to analyze association of mTOR with raptor and rictor. Immunoprecipitates and total cell lysates were analyzed by immunoblotting (5 ug input). β-Actin served as a loading control. Data are representative of at least three independent experiments.