FIGURE 2:

PI3K-C2α is required for mTORC1 activation and translocation. (A) siRNA directed against PI3K-C2α was electroporated into 3T3-L1 adipocytes for 96 h. Cells were serum-deprived for 3 h, stimulated with 100 nM insulin for 10 min, and blotted as indicated. (B) 3T3-L1 adipocytes were electroporated with control or PI3K-C2α siRNA for 96 h. Cells were stimulated as described, fixed, and then stained with anti-mTOR antibodies. Scale bars: 10 μm. (C) Cells were quantified by counting the percentage of cells in which plasma membrane staining of mTOR was visible. These data represent three independent experiments in which >150 cells were counted. Asterisks indicate a statistically significant difference (p < 0.05).