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. 2014 Oct 23;16(10):824–834. doi: 10.1016/j.neo.2014.08.006

Figure 4.

Figure 4

Effect of S6K1 knockdown on feedback PI3K pathway activation and cell proliferation. (A) S6K1 knockdown. A375 cells seeded in a six-well plate were transfected with scrambled or S6K1 siRNA (2.5 nmol each). After incubation for 48 hours, the cells were harvested and analyzed for S6K1 expression and phosphorylation of the indicated proteins by Western blot. (B) The effect of S6K1 knockdown on cell proliferation. A375 cells seeded in a 96-well plate were transfected with a scrambled control siRNA or S6K1 siRNA. After incubation overnight, the cells were incubated in the absence or presence of A77 1726 (100 or 200 μM) for 72 hours and analyzed for cell proliferation by an ATP-based Cell-Glo assay. Data represent the mean ± SD from one of two experiments in triplicate with similar results. *P < .05, compared to control siRNA-transfected A375 cells.