Depletion of ATG5 and overexpression of USP30 rescue pexophagy in UBXD8 KO cells. (A) Representative images of HeLa cells transfected with control, UBXD8, and ATG5 siRNAs and stained for catalase. (B) Quantification of peroxisomes per cell from A. 50–100 cells were analyzed in N = 3 independent experiments. The violin plot shows median and 95% confidence intervals. ****P < 0.0001, two-way ANOVA with Dunnett’s multiple comparisons test. (C) Immunoblot of UBXD8 and ATG5 depletion. (D) Representative images of HeLa cells transfected with control, UBXD8, or NBR1 siRNAs and stained for catalase. (E) Quantification of peroxisomes per cell from D. 50–100 cells were analyzed in N = 3 independent experiments. Violin plot shows median and 95% confidence intervals. ****P < 0.0001, two-way ANOVA with Dunnett’s multiple comparisons test. (F) Immunoblot of UBXD8 and NBR1 depletion. (G) WT and UBXD8 KO HeLa cells were stained with NBR1 and catalase. (H) Mander’s colocalization of images in G. 100–120 cells were analyzed in N = 3 independent experiments. The violin plot shows median and 95% confidence intervals. ****P < 0.0001, unpaired t test. The scale bar is 10 μM (A and D) and 5 μM (D and G). Source data are available for this figure: SourceData F8.