mTORC1, but not mTORC2, is necessary for myeloid differentiation of mEB8-ER cells. (A,C,E) Top panel: Western blot of mTOR (A), Raptor (C), or S6K1 (E) in mEB8-ER cells with or without shRNAs targeting mTOR, Raptor, or S6K1. α-tubulin was a loading control. A typical blot from 4 independent experiments. Bottom panel: Quantification of blots from 4 separate experiments. Each bar represents the mean ± SEM. *P < .05. LMP/pMKO: the empty retrovector. (B,D,F) Relative percentage of CD11b and Gr-1–positive cells with or without mTOR (B), Raptor (D), and S6K1 (F) depletion, measured with flow cytometry. Four separate experiments were conducted, and quantification of 3 replicates of a typical experiment. Each bar represents the mean ± SEM (error bars). All values were normalized to the level (= 1) in cells without depletion. *P < .05.