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. 2012 Feb 3;31(5):1134–1146. doi: 10.1038/emboj.2011.478

Figure 6.

Figure 6

Inhibition of mTORC1 activity inhibits exit from contact inhibition of Tsc2−/− cells but not of 4E-BP1/2 DKO cells. (A, B) Two days post confluency, Tsc2+/−, Tsc2−/− MEFs (A) and 4E-BP WT, 4E-BP1/2 DKO MEFs (B) were incubated with rapamycin (20 nM) 3 h prior to being induced with 20% FBS. Cells were then harvested and counted at 0, 24, 48, and 72 h post stimulation. The relative increase in cell number (compared with time 0) is depicted. Results represent the average ±s.e. of three independent experiments. (C, D) Cell lysates of cells treated as in (A) and (B) were prepared at 0, 16, and 24 h post addition of 20% FBS, and subjected to immunoblotting with anti-phospho-S6K, S6K, 4E-BP1, phospho-H3, Skp2, and tubulin antibodies.