Various cellular stressors induce TFEB activation in either an ATG conjugation system–independent or an ATG conjugation system–dependent manner. (A) Representative fluorescence images of WT, ATG7 KO, and ATG13 KO HeLa cells stably expressing TFEB::mNG and Myc::Parkin, either untreated (control) or treated with O/A for 3 h. (B) Quantification of the image data shown in A (n = 3 biologically independent samples). (C) Representative fluorescence images of WT, ATG7 KO, and FIP200 KO HeLa cells stably expressing TFEB::mNG, either untreated (control) or treated with etoposide for 24 h. (D) Quantification of the image data shown in C (n = 3 biologically independent samples). (E) Representative fluorescence images of WT, ATG7 KO, and FIP200 KO HeLa cells stably expressing TFEB::mNG, either untreated (control) or treated with arsenite for 6 h. (F) Quantification of the image data shown in E (n = 3 biologically independent samples). (G) Representative fluorescence images of WT, ATG7 KO, and FIP200 KO HeLa cells stably expressing TFEB::mNG, either untreated (control) or treated with MG132 for 9 h. (H) Quantification of image data shown in G (n = 3 biologically independent samples). (I) Representative immunoblots of LC3B and GAPDH in ATG13 KO HeLa cells stably expressing Myc::Parkin or in FIP200 KO HeLa cells. Cells were either untreated (control) or treated with the indicated reagents. (J) Quantification of the image data shown in I (n = 3 biologically independent samples). The value of “Control” was defined as 1, and relative values compared with each control were calculated. (A, C, E, and G) Scale bars, 50 µm. (A, C, E, and G) Arrowheads indicate nuclear-localized TFEB::mNG. (B, D, F, H, and J) P values were determined by ANOVA with Tukey’s multiple comparison test; *P < 0.05, **P < 0.01, ***P < 0.005. Source data are available for this figure: SourceData F4.
Figure 4.

Various cellular stressors induce TFEB activation in either an ATG conjugation system–independent or an ATG conjugation system–dependent manner. (A) Representative fluorescence images of WT, ATG7 KO, and ATG13 KO HeLa cells stably expressing TFEB::mNG and Myc::Parkin, either untreated (control) or treated with O/A for 3 h. (B) Quantification of the image data shown in A (n = 3 biologically independent samples). (C) Representative fluorescence images of WT, ATG7 KO, and FIP200 KO HeLa cells stably expressing TFEB::mNG, either untreated (control) or treated with etoposide for 24 h. (D) Quantification of the image data shown in C (n = 3 biologically independent samples). (E) Representative fluorescence images of WT, ATG7 KO, and FIP200 KO HeLa cells stably expressing TFEB::mNG, either untreated (control) or treated with arsenite for 6 h. (F) Quantification of the image data shown in E (n = 3 biologically independent samples). (G) Representative fluorescence images of WT, ATG7 KO, and FIP200 KO HeLa cells stably expressing TFEB::mNG, either untreated (control) or treated with MG132 for 9 h. (H) Quantification of image data shown in G (n = 3 biologically independent samples). (I) Representative immunoblots of LC3B and GAPDH in ATG13 KO HeLa cells stably expressing Myc::Parkin or in FIP200 KO HeLa cells. Cells were either untreated (control) or treated with the indicated reagents. (J) Quantification of the image data shown in I (n = 3 biologically independent samples). The value of “Control” was defined as 1, and relative values compared with each control were calculated. (A, C, E, and G) Scale bars, 50 µm. (A, C, E, and G) Arrowheads indicate nuclear-localized TFEB::mNG. (B, D, F, H, and J) P values were determined by ANOVA with Tukey’s multiple comparison test; *P < 0.05, **P < 0.01, ***P < 0.005. Source data are available for this figure: SourceData F4.

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