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. 2010 Sep 17;285(47):36387–36394. doi: 10.1074/jbc.M110.169284

FIGURE 3.

FIGURE 3.

RSV-promoted Inhibition of mTOR activation is TSC1/2- and AMPK-independent. A, serum-starved TSC1/2+/+ and TSC1/2−/− MEF cells were pretreated with 10 mm leucine for 60 min and then cotreated with or without 50 μm RSV for 30 min. B, serum-starved TSC1/2+/+ and TSC1/2−/− MEFs were pretreated with 50 μm RSV for 20 min, followed with or without 10 nm insulin (Ins) for 10 min. KO, knock-out. C, serum-starved AMPKα2-suppressed C2C12 cells were pretreated with or without leucine for 60 min and then cotreated with or without 50 μm RSV for 30 min. For all experiments, the phosphorylation and protein levels of S6K, 4E-BP1, and AMPK and the protein levels of TSC1 and TSC2 in cell lysates were determined by Western blotting with the indicated antibodies. All data are representative of at least three independent experiments with similar results, and A–C were quantified using the NIH Scion Image program. Differences between groups were examined for statistical significance using ANOVA. *, p < 0.05; **, p < 0.01. N, no addition.