Extended Data Fig. 10: Genetic targeting of Pla2g15 ameliorates defects in NPC1-deficient mouse.
From: PLA2G15 is a BMP hydrolase and its targeting ameliorates lysosomal disease
a, PLA2G15 depletion increases levels of only few BMPs in NPC1-deficient mice. Fold changes in the levels of selected BMPs measured on the QQQ mass spectrometer in all genotypes over Npc1+/+ Pla2g15+/+ brain (left) and liver (right). Data are mean ± s.d. (n = 4, for all genotypes except n = 3 for Npc1+/+ Pla2g15-/-). Statistical analysis was performed using two-tailed unpaired t-tests.*p < 0.05 and **p = 0.0072. All measured BMP species and their statistics are presented in Supplementary Table 7. b, Cholesterol staining. Cholesterol was measured in left hemibrain and left liver lobe (n = 4 per group). Data are mean ± s.d. Statistical analysis was performed using one-way ANOVA followed by Bonferroni’s multiple comparisons. **p = 0.00295. c,d, Genetic depletion of PLA2G15 reverses secondary lipid storage in NPC1-deficient brain (c) and liver (d) mouse tissues. Fold changes in the levels of selected total lipid classes in all genotypes compared to Npc1+/+ Pla2g15+/+ brain (c) and liver (d). Data are mean ± s.d. (n = 5, for all brain genotypes except n = 6 for brain Npc1-/- Pla2g15-/- and n = 6, for all liver genotypes). Statistical analysis was performed using two-tailed unpaired t-tests.*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. All measured individual lipid species, total lipid classes and their statistics are presented in Supplementary Table 8. e,f, Purkinje cell count by histomorphometry on Calbindin-immunolabelled CNS sections. e, Proof-of-concept of the image analysis indicating the annotations in the cerebellum (ROI: Region of Interest) and the detection of Purkinje cell soma at different magnifications. f, Representative images of the cerebellum at the level of lobe II/III for comparison among study groups. No label: Scanned whole slide images (WSI) prior to image analysis. Labeled: WSI indicating the annotations of the ROI and the detections of Purkinje cell’s soma. g,h, Genetic ablation of PLA2G15 decreases histopathology lesions in NPC1-deficient mice. The microscopy images are in g and their mean severity scores of each histopathology finding are shown in h. Data are mean ± s.d of n = 6 independent replicates. **p = 0.0063, ***p = 0.0006 and ****p < 0.0001, by one-way ANOVA Fisher’s LSD test comparing NPC1-deficient tissues to other genotypes. Scale bar is 20 µm. i, Body weight assessment for all genotypes. Animal body weight was measured every other day starting at 3 weeks old. Data are mean ± s.e.m. of n = 12, 6 males and 6 females per genotype.
