NVP-BEZ235 targets both rictor and raptor. (A) Thymidine uptake assay. BCWM.1 cells were cultured with rapamycin and LY294002 for 48 hours. (B-C) BCWM.1 cells were cultured with NVP-BEZ235 (20nM), rapamycin (5nM), LY294002 (10μM), or the combination of rapamycin and LY294002 for 6 hours. (B) Whole-cell lysates were subjected to immunoprecipitation (IP) using a mTOR antibody and Western blotting using anti-mTOR, anti-rictor, and anti-raptor antibodies. (C) Whole-cell lysates were subjected to Western blot using anti–p-Akt, anti-Akt, anti–p-p70S6, anti–p-4EBP1, anti–p-ERK, anti-ERK, and anti–α-tubulin antibodies.