Mito-VPS41 induces redistribution of LAMP1 to mitochondria. (A) Schematic representation of the experimental setup. LAMP1-positive membranes recruited by mito-VPS41 could represent LAMP carriers and/or late endosomes/lysosomes. (B) Confocal immunofluorescence distribution of endogenous LAMP1 and negative control mito-V5, mito-VPS41^WT-V5, or mito-VPS41^R662*-V5 (stained with anti-V5) expressed in HeLa^VPS41KO cells. The bottom two rows show confocal immunofluorescence images of endogenous LAMP1 and mito-VPS18-GFP or mito-VPS39-GFP in HeLa^VPS18KO and HeLa^VPS39KO cells, respectively. Dashed boxes are enlarged in the next images. (C) Scatter plot quantification of LAMP1 recruitment to mitochondria upon the expression of mito-V5, mito-VPS41^WT-V5, mito-VPS41^R662*-V5, mito-VPS18-GFP, or mito-VPS39-GFP as shown in B. Scatter plot data are presented as the mean ± SEMreen dots for experimental replicate means. ****P < 0.0001, ***P < 0.001, ns: not significant. n ≥ 25. Statistical significance was obtained using Tukey’s multiple comparisons test. Scale bars: 20 or 5 µm (inset magnification).