|
Fig. 5 Patient-derived fibroblasts accumulate apoptotic material in the gastrosome that is larger in NPC deficient cases.a Representative images of human patients? fibroblasts stained for LAMP1. The gastrosome is indicated with an orange arrowhead. Scale bar 10 µm. b Representative image of HeLa-fed human patients? fibroblasts stained for Cathepsin D. The gastrosome is indicated with an orange arrowhead. Scale bar 10 µm. c Diameter of the gastrosome in HeLa-fed human patients? fibroblasts: HI (n = 114), HI treated with HP?CD (n = 113), HI treated with U18666a (n = 126), HI treated with U18666a and HP?CD (n = 176), NPC-P1 (n = 156), NPC-P1 treated with HP?CD (n = 146), NPC-P2 (n = 113), NPC-P2 treated with HP?CD (n = 94). d Representative EM image of the gastrosome in HeLa-fed human patients? fibroblast. Scale bar 3 µm. e Representative images of HeLa-fed human patients? fibroblast stained with filipin. The gastrosome is indicated with an orange arrowhead and lysosomal pools are framed in blue squares. Scale bar 10 µm. f, g Mean intensities of the filipin staining from HeLa-fed human patients? fibroblasts: HI (n = 25), HI treated with HP?CD (n = 30), HI treated with U18666a (n = 16), HI treated with U18666a and HP?CD (n = 32), NPC-P1 (n = 21), NPC-P1 treated with HP?CD (n = 21), NPC-P2 (n = 23), NPC-P2 treated with HP?CD (n = 33) in the lysosomal pool (framed in blue squares in Fig. 5e) (f), in the gastrosome (indicated with an orange arrowhead in Fig. 5e) (g). n refers to the number of cells examined. Boxplots represent the median value and interquartile range; the ends of the whiskers correspond to the minimum and maximum values. Statistical tests: Mann-Whitney-Wilcoxon test two-sided with Bonferroni correction. ns without any additional p-value on the graph stands for p = 1. HI stands for healthy individual. NPC1-P1 corresponds to GM18393 NPC patient and NPC1-P2 corresponds to GM17919 NPC patient. Source data are provided as a Source Data file.