Fig. 2

Quantitative Western blot analysis of TFEB protein levels in response to mTOR activity modulations. a HeLa cells were treated with fresh FM to enhance mTOR activity. At the indicated time points, levels of TFEB and phosphorylated 4E-BP1 (p-4E-BP1) were analyzed by Western blotting. Lanes for time point ‘0’ originate from same membrane as later time points. b Quantified values for TFEB, normalized to loading control GAPDH, shown relative to time point ‘0’. c Quantified values for p-4E-BP1, normalized to total 4E-BP1, shown relative to time point ‘0’. d HeLa cells were treated with FM containing 2 μM Torin1. At the indicated time points, levels of TFEB and phosphorylated 4E-BP1 (p-4E-BP1) were analyzed by Western blotting. e Quantified values for TFEB, normalized to loading control GAPDH, shown relative to time point ‘0’. f Quantified values for p-4E-BP1, normalized to total 4E-BP1, shown relative to time point ‘0’. Error bars denote mean ± SD of three independent experiments. Statistical significances were tested vs. time point ‘0’ (Student’s two-tailed t-test; *, p ≤ 0.05; ***, p ≤ 0.001)