FIGURE 5: AZD induces TFEB/TFE3 activation. (A-F) TFEB-GFP-expressing or wild type U2OS cells were treated with Torin 1 (300 nM) or AZD (40 µM) combined or not with NAC (3 mM) for 6 h. The average nuclear intensity of TFEB was measured (A, B), TFE3 translocation was assessed by immunostaining (C, D), and (E) TFEB and (F) TFE3 in the cytoplasm and nucleus were assessed by Western blot. (G) Western blot was performed for measuring the expression of TFEB and TFE3 in U2OS cells with or without TFEB-/-TFE3-/- double-knockout treated with AZD. Scale bars equal 10 μm. (*P < 0.05, **P < 0.01, ***P < 0.001,****P<0.0001 vs. DMSO; #P < 0.05, ##P < 0.01, ###P < 0.001, #### P < 0.0001 vs. Cont or WT).