Fig. 3

Localization of VPS13C at the late endosomes and lysosomes disturbed by p.Trp395Cys and p.Ala444Pro. Triple immunofluorescence of V5 (VPS13C constructs, green) with markers for late endosomes (Rab7) and lysosomes (Lamp1) in HeLa cells. a Cells expressing VPS13C WT showed a clear vesicular staining pattern while those expressing the missense mutations p.Trp395Cys or p.Ala444Pro mislocalized in larger cytosolic structures. b Quantification of cells that appeared either vesicular or mislocalized. c Staining with anti-V5 antibody in WT-overexpressing HeLa cells showed co-staining with markers against late endosomes and lysosomes. HeLa cells overexpressing either p.Trp395Cys or p.Ala444Pro showed no co-localization with endosomes and lysosomes. White lines indicate the position of the intensity profiles. Scale bars = 5 µm