Increased lysosomal tethering affects lysosomal motility and distribution in VPS13C-deficient dopaminergic neurons. (A and B) Representative live-cell confocal images from control (A; Video 1) and VPS13C KD (B; Video 2) neurons showing LAMP1-positive vesicles (green, upper panel) and lysosomal motility demonstrated by kymographs (bottom panels). Dashed line represents the outline of the cell. Scale bar: 10 µm, kymograph scale bar: x = 1 µm, y = 20 s. (C–E) Quantification of the average distance traveled by lysosomes in 60 s and quantification of the average lysosomal motility with histogram distribution. N = 4, from n = 11 cells (control) and n = 13 cells (VPS13C KD). (F) Histogram distribution of the average lysosomal distance to the nucleus. N = 4, from n = 15 cells (control) and n = 18 cells (VPS13C KD). (G) Quantification of the percentage of lysosomes in the perinuclear region (<7 µm distance from nuclear membrane). (H) Quantification of distal lysosomes (≥7 µm distance from nuclear membrane). Data represented as mean ± SEM; unpaired two-tailed t test (C, D, G, and H); *P < 0.05 (D, G, and H), **P < 0.01 (C).