Fig. 1.

TFEB is subject to nuclear export. a Immunofluorescence with indicated antibodies using control MCF7 cells or those treated with Torin 1 (250 nM, 1 h). n > 30 cells per condition. b Real time imaging of MCF7 stable cell lines expressing doxycycline-inducible TFEB-GFP (iTFEB-GFP) imaged at indicated intervals in the absence of doxycycline. Images derived from Supplementary Movie 1. c Live cell imaging of MCF7 iTFEB-GFP cells before and after 1 h Torin 1 (250 nM) or 2 h U0126 (20 μM) treatment or after removal of the drug (upper panels) as indicated, or 30′ and 1 h after removal of Torin 1 (lower panels). n > 30 cells per condition. d Imaging of MCF7 iTFEB-GFP or iGFP cells before and after LMB (2 h; 20 nM) treatment. Quantification is derived from one experiment that is representative of 2 independent experiments. n > 30 cells per condition. e Immunofluorescence using MCF7 or HT29 cells and indicated antibodies with or without treatment with Torin 1 (1 h; 250 nM), LMB (2 h; 20 nM), and U0126 (3 h; 20 μM). n > 500 cells per condition. Details of all quantifications including number of replicates and cells imaged are provided in Supplementary Table 1 and Methods. Scale bars = 20 μM. Error bars = SD. Student’s t-test; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, n.s. not significant