Figure 2.

mTORC1 is localized at FAs. (A and B) HeLa cells grown in full-nutrient medium were FCS starved for 18 h (−FCS) or FCS starved and then recovered in FCS-containing medium for 10 min (10 min FCS) and immunostained for mTOR (A) or LAMP1 (B) and the FA protein paxillin. Colocalization (Manders coefficient) was analyzed. (C) PLAs for mTOR–paxillin interactions (and SDHA–paxillin as a control) were performed in HeLa cells under starvation–refeeding conditions as in A and B. Cells were counterstained with paxillin antibody to mark FAs, and the proportions of PLA-positive cells and PLA dots/cell were quantified. Error bars represent SEM; n = 3 independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001; two-sided Student’s t test. Scale bars, 20 µm (insets, 10 µm). Nuclei were visualized with DAPI.