Loss of VPS13C disrupts lysosomal morphology in hiPSC-derived dopaminergic neurons. (A and B) Representative immunoblot and quantification of VPS13C KD efficiency in hiPSC-derived dopaminergic neurons (day 70) after 14 days of treatment with control and VPS13C shRNA. (C) Representative live-cell confocal images of LAMP1-mGFP (green) -positive vesicles in control (upper) and VPS13C KD (lower) neurons with insets showing smaller lysosomes (white arrow) in control condition versus enlarged lysosomes (yellow arrow) in VPS13C KD condition. Dashed line represents the outline of the cell (scale bar: 10 µm, inset: 1 µm). (D) Quantification of the percentage of cell area occupied by lysosomes from LAMP1-mGFP live-cell confocal imaging (N = 4, from n = 15 cells). (E and F) Quantification and histogram distribution of average lysosomal size in control and VPS13C KD dopaminergic neurons (N = 4, from n = 15 cells). (G) Representative images of PFA-fixed dopaminergic neurons with LAMP1-mGFP expression (green) and co-staining with TH (magenta) and DAPI (blue), with insets showing smaller lysosomes in control condition (upper panel, white arrow) and enlarged clustered lysosomes in VPS13C KD neurons (lower panel, yellow arrow) (scale bar: 10 µm, inset: 1 µm). Data are represented as mean ± SEM; unpaired two-tailed t test (B, D, and E); *P < 0.05 (D), ****P < 0.0001 (B and E). Source data are available for this figure: SourceData F1.