Fig. 6: USP13 promotes the mitochondria translocation of STAT1 and improves mitochondrial complex I activity and ATP synthesis.
a–d, j, k HL-1 were transfected with plasmids of empty vector (EV) or USP13 (USP13OE) followed by Ang II stimulation (1 μM, 24 h). HL-1 were pretreated with Spautin-1 (USP13i, 10 μM, 1 h) followed by Ang II stimulation. a Levels of P-STAT1 and STAT1 in mitochondria was detected by western blot. Tom 20 was used as loading control. b The P-STAT1 mitochondria translocation was detected by immunofluorescence. c, d Representative images of JC-1 staining (c; green: JC-1 monomers meaning decreased MMP; red: JC-1 aggregates meaning increased MMP) and the quantitative analysis (d; n = 6 independent experiments, adjusted P values were determined by one-way ANOVA with Bonferroni’s correction; black column: Ang II + EV, purple column: Ang II+Veh). e GSEA enrichment analysis of transcriptome of heart tissues from Ang II- induced USP13cKO mice and USP13f/f mice (NES: normalized enrichment score; FDR: false discovery rate). f–i Complex I activity and ATP levels were detected in heart tissues from Ang II- (f, g) or TAC- (h, i) induced USP13f/f mice and USP13cKO mice (n = 6, adjusted P values were determined by one-way ANOVA with Bonferroni’s correction). j, k Complex I activity (j) and ATP levels (k) were detected in HL-1 (n = 6 independent experiments, adjusted P values were determined by one-way ANOVA with Bonferroni’s correction; black column: Ang II + EV, purple column: Ang II+Veh). l, m STAT1 knockdown HL-1 cells were transfected with plasmids of EV or USP13OE followed by Ang II. Complex I activity (l) and ATP levels (m) were detected (n = 6 independent experiments, adjusted P values were determined by one-way ANOVA with Bonferroni’s correction). For (d, f–m), data are presented as mean ± s.e.m.
