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. Author manuscript; available in PMC: 2016 Jul 16.
Published in final edited form as: Cell. 2015 Jul 2;162(2):259–270. doi: 10.1016/j.cell.2015.06.017

Figure 5. mTORC1 Signaling Is a Negative Regulator of Extracellular Protein-Dependent Growth.

Figure 5

(A) Cell numbers of K-RasG12D MEFs at day 3 of culture in leucine-containing or leucine-free medium + 3% albumin and following inhibitors: MEK1/2 (1 μM PD0325901, 50 μM PD98059), PI3-kinase (25 μM LY294002, 2 μM wortmannin), tyrosine kinases (50 μM genistein), mTOR (50 nM rapamycin, 250 nM torin 1), mTOR/PI3-kinase (0.5 μM BEZ235, 0.5 μM GDC0980). Dashed lines indicate starting cell numbers. (B) mTOR, PI3-kinase and MAP kinase pathway activity in K-RasG12D MEFs cultured for 1 day in leucine-free medium + 3% albumin, analyzed by WB. Inhibitors were as in (A) (C) Growth curve of K-RasG12D MEFs in leucine-free medium ± 3% albumin and indicated concentrations of torin 1. (D) Growth curve of K-RasG12D MEFs expressing shRNA against Raptor, Rictor or control in leucine-free medium ± 3% albumin. (E) Bright field images of K-RasG12D MEFs expressing shRNA against Raptor, Rictor or control at day 4 of culture in leucine-free medium ± 3% albumin. Scale bars = 50 μm.

Data are represented as mean ± STDEV (n = 3). See also Fig. S5.