Depletion of NBR1 and MARCH5 rescues pexophagy in UBXD8 null cells. (A) Representative images of HeLa cells transfected with control, UBXD8, and MARCH5 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 MARCH5 depletion. (D) Representative images of HeLa cells (WT and UBXD8 KO) transfected with GFP-USP30. Cells were stained for catalase. (E) Quantification of peroxisomes per cell in GFP-USP30–transfected cells. 50–100 cells were analyzed in N = 3 independent experiments. The violin plot shows median and 95% confidence intervals. ***P < 0.001, ****P < 0.0001, two-way ANOVA with Dunnett’s multiple comparisons test. (F) Immunoblot of GFP-USP30 expression. The scale bar is 10 μM (A and D). Source data are available for this figure: SourceData F9.