NPC1 deficiency causes expansion of the gastrosome in phagocytic cells.a Representative images of OT microglia (TgBAC(csf1ra:GAL4-VP16), Tg(UAS-E1B:NTR-mCherry)) in the zebrafish displayed as maximum projections and segmented images. The largest vesicle present in each cell is displayed as a segmentation. Scale bar 5 µm. b Diameter of the largest vesicle in wild-type (N = 26, n = 102), NPC1 CRISPANT (N = 29, n = 232) and NPC2 CRISPANT (N = 7, n = 126) OT microglia. N refers to the number of zebrafish embryos and n to the number of microglia examined. c Graph representing the diameter of the largest vesicle found in NPC1 CRISPANT OT microglia over time. Colored vertical lines indicate fusion with incoming phagosomes. d Representative images of fusion events between the large vesicle (marked with red) and incoming phagosome (marked with green) in NPC1 CRISPANT OT microglia (TgBAC(csf1ra:GAL4-VP16), Tg(UAS-E1B:NTR-mCherry)). Scale bar 10 µm. Related to Supplementary Movie 3. e Representative images of RAW LAMP1-EGFP macrophages. Orange arrowhead indicates the gastrosome. Scale bar 10 µm. f Diameter of the gastrosome in HeLa-fed RAW LAMP1-EGFP macrophages, untreated (n = 69) and treated with U18666a (n = 115). n refers to the number of cells examined. g EM image of HeLa-fed RAW treated with U18666a. Lysosomes are framed in colored squares and the gastrosome is framed in the black square (enlargement in Fig. 3h). Scale bar 5 µm. Enlarged regions with lysosomes on the right. Scale bar 0.25 µm. h Electron microscope image of the gastrosome in HeLa-fed RAW treated with U18666a. Scale bar 3 µm. Statistical tests: Mann-Whitney-Wilcoxon test two-sided with Bonferroni correction. OT stands for optic tectum. Source data are provided as a Source Data file.
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