Figure 4.

Mitophagy induction in response to the mitochondrial recruitment of ATG16L1[100-250] or ULK1 is blocked in RB1CC1 KO cells. (A) WT, RB1CC1 KO #52 or RB1CC1 KO #118 ARPE-19 mito-QC cells were transduced to express the indicated proteins or not transduced (CTRL) for 48 h before cell lysis and immunoblot analysis. Left: Representative immunoblot. Right: Quantification of mitophagy showing the levels of the mitochondrial proteins HSPD1 and OPA1 normalized to TUBA from a minimum of 5 independent experiments. Statistics: Two-way ANOVA + Tukey’s multiple comparisons test. (B) WT, RB1CC1 KO #52 or RB1CC1 KO #118 ARPE-19 mito-QC cells were transduced to express the indicated proteins (or not transduced [CTRL]) for 48 h before microscopy analysis. Left: Representative images. Scale bar: 10 µm. Right: Quantification of mitophagy showing the mean number of mitolysosomes per cell from 4 independent experiments with a minimum of 30 cells being analyzed per condition in each experiment. Statistics: Two-way ANOVA + Tukey’s multiple comparisons test.