TFEB activation by several cellular stressors involves either Mode I or Mode II. (A) Representative fluorescence images of ATG7 KO HeLa cells stably expressing TFEB::mNG and Myc::Parkin, either untreated (control) or treated with O/A for 3 h. Cells were transfected with siLuc, siCCT7, or siTRIP6. (B) Quantification of the image data shown in A (n = 3 biologically independent samples). (C) Representative fluorescence images of WT or ATG7 KO HeLa cells transiently expressing mNG::TRIP6, either untreated (control) or treated with O/A for 3 h. (D) Representative fluorescence images of ATG7 KO HeLa cells stably expressing TFEB::mNG, either untreated (control) or treated with etoposide for 24 h. Cells were transfected with siLuc, siCCT7, or siTRIP6. (E) Quantification of the image data shown in D (n = 3 biologically independent samples). (F) Representative fluorescence images of WT or ATG7 KO HeLa cells transiently expressing mNG::TRIP6, either untreated (control) or treated with etoposide for 24 h. (G) Representative fluorescence images of ATG7 KO HeLa cells stably expressing TFEB::mNG, either untreated (control) or treated with MK6-83 for 3 h. Cells were transfected with siLuc, siCCT7, or siTRIP6. (H) Quantification of the image data shown in G (n = 3 biologically independent samples). (I) Representative fluorescence images of WT or ATG7 KO HeLa cells transiently expressing mNG::CCT7, either untreated (control) or treated with MK6-83 for 3 h. (J) Representative fluorescence images of WT HeLa cells stably expressing TFEB::mNG, either untreated (control) or treated with arsenite for 6 h or MG132 for 9 h. Cells were transfected with siLuc or siAPEX1. (K) Quantification of image data shown in J (n = 3 biologically independent samples). (A, C, D, F, G, I, and J) Scale bars, 50 µm. (A, D, and G) Arrowheads indicate nuclear-localized TFEB::mNG. (C, F, and I) Arrowheads indicate TRIP6/CCT7 dots. (B, E, H, and K) Relative value of the TFEB nuclear translocation rate under drug treatment conditions compared with that under basal (control) conditions. P values were determined by ANOVA with Tukey’s multiple comparison test (B, E, and H) or t test (K); *P < 0.05, **P < 0.01.
Figure 5.

TFEB activation by several cellular stressors involves either Mode I or Mode II. (A) Representative fluorescence images of ATG7 KO HeLa cells stably expressing TFEB::mNG and Myc::Parkin, either untreated (control) or treated with O/A for 3 h. Cells were transfected with siLuc, siCCT7, or siTRIP6. (B) Quantification of the image data shown in A (n = 3 biologically independent samples). (C) Representative fluorescence images of WT or ATG7 KO HeLa cells transiently expressing mNG::TRIP6, either untreated (control) or treated with O/A for 3 h. (D) Representative fluorescence images of ATG7 KO HeLa cells stably expressing TFEB::mNG, either untreated (control) or treated with etoposide for 24 h. Cells were transfected with siLuc, siCCT7, or siTRIP6. (E) Quantification of the image data shown in D (n = 3 biologically independent samples). (F) Representative fluorescence images of WT or ATG7 KO HeLa cells transiently expressing mNG::TRIP6, either untreated (control) or treated with etoposide for 24 h. (G) Representative fluorescence images of ATG7 KO HeLa cells stably expressing TFEB::mNG, either untreated (control) or treated with MK6-83 for 3 h. Cells were transfected with siLuc, siCCT7, or siTRIP6. (H) Quantification of the image data shown in G (n = 3 biologically independent samples). (I) Representative fluorescence images of WT or ATG7 KO HeLa cells transiently expressing mNG::CCT7, either untreated (control) or treated with MK6-83 for 3 h. (J) Representative fluorescence images of WT HeLa cells stably expressing TFEB::mNG, either untreated (control) or treated with arsenite for 6 h or MG132 for 9 h. Cells were transfected with siLuc or siAPEX1. (K) Quantification of image data shown in J (n = 3 biologically independent samples). (A, C, D, F, G, I, and J) Scale bars, 50 µm. (A, D, and G) Arrowheads indicate nuclear-localized TFEB::mNG. (C, F, and I) Arrowheads indicate TRIP6/CCT7 dots. (B, E, H, and K) Relative value of the TFEB nuclear translocation rate under drug treatment conditions compared with that under basal (control) conditions. P values were determined by ANOVA with Tukey’s multiple comparison test (B, E, and H) or t test (K); *P < 0.05, **P < 0.01.

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