Skip to main content

Advertisement

Fig. 1 | BMC Cancer

Fig. 1

From: Time course decomposition of cell heterogeneity in TFEB signaling states reveals homeostatic mechanisms restricting the magnitude and duration of TFEB responses to mTOR activity modulation

Fig. 1

Characterization of the effect of Torin1 and fresh nutrients on mTOR and endogenous TFEB. a Schematic representation of the effects of Torin1 and fresh fully-supplemented medium (FM) on the regulation of TFEB by mTOR. b Dose-response of the effect of Torin1 on mTOR activity. HeLa cells were treated with FM containing the indicated concentrations of Torin1, or kept in culture medium (non-treated, NT), for 1.5 or 3 hours, and phosphorylation of the mTOR substrates 4E-BP1 and p70-S6K1 was measured by Western blotting. c Quantification of the ratio of phosphorylated 4E-BP1 (p-4E-BP1) to total 4E-BP1. Graphs display mean values of three independent experiments normalized to NT values. Error bars denote mean ± SD of three independent experiments. Statistical significance was tested vs. NT conditions (Student’s two-tailed t-test; **, p ≤ 0.01; ***, p ≤ 0.001). d Immunofluorescence of TFEB and p-4E-BP1, as a read-out for mTOR activity, in response to FM and Torin1. HeLa cells were kept in culture medium (non-treated, NT), treated with fresh FM, or with FM supplemented with Torin1 (2 μM) for 3 hours and immunostained for TFEB and p-4E-BP1. To reveal varying intensity levels the look-up-table ‘Fire’ (ImageJ) was applied to grey scale images, representing intensity values ranging from low (dark purple) to high (white) as displayed in color scale bar. Scale bars, 20 μm

Back to article page