Regulatory mechanism and downstream targets of TFEB during lysosomal damage. (A and B) Representative immunoblots of TFEB, S6K, p-S6K, and GAPDH in WT or ATG7 KO HeLa cells treated with 1 mM LLOMe (A) or 100 μM MK6-83 (B) for the indicated times. (C and D) Quantification of image data of pS6K/S6K shown in A and B (n = 3 biologically independent samples). (E) Venn diagram showing the overlap between two DEGs: 471 genes were TFEB-dependent targets during starvation (pink circle), while 147 genes were TFEB-dependent targets under LLOMe treatment (green circle). Only three genes were regulated under both starvation and LLOMe treatment. (F and G) Quantification of the relative expression of IL-6, CCL21 by qPCR in WT, ATG7 KO, or TFEB KO HeLa cells under nontreated(control) or 1 mM LLOMe-treated conditions (n = 3 biologically independent samples). (C, D, F, and G) P values were determined by ANOVA with Tukey’s multiple comparison test; *P < 0.05, ****P < 0.0001. Source data are available for this figure: SourceData FS1.