Extended Data Fig. 8: PLA2G15 role in lysosomal lipid metabolism.
From: PLA2G15 is a BMP hydrolase and its targeting ameliorates lysosomal disease
a,b, PLA2G15 supplementation reduces GCase activity in bone marrow derived macrophages (BMDMs). a, Representative fluorescence microscopy images of BMDMs treated with 300 nM recombinant PLA2G15 WT, inactive PLA2G15 S198A or vehicle buffer (PBS) overnight followed by a 30-min incubation of 10 µM LysoFQ-GBA to measure GCase activity. b, Quantification of signal intensity of LysoFQ-GBA. Signals were normalized to vehicle treated BMDMs. Data are mean ± s.e.m. (Vehicle n = 194 fields, PLA2G15 WT n = 118 and PLA2G15 S198A n = 83 of cells). *p < 0.05, by one-way ANOVA Fisher’s LSD test. c, Pla2g15 knockdown in wildtype and mutant Gba mouse-derived BMDMs after two-day treatment with 10 nM control siRNA or two different siRNA that target Pla2g15. qPCR quantification of Pla2g15 mRNA abundance relative to siControl. Actin was used an endogenous control. Data are mean ± s.e.m. (n = 6 per condition). ****p < 0.0001, by one-way ANOVA Fisher’s LSD test. d, PLA2G15 knockdown in two independent NPC1 patient fibroblast cell lines (Coriell GM03123 and GM18453) after two-day treatment with 10 nM control siRNA or two different siRNA that target PLA2G15. qPCR quantification of PLA2G15 mRNA abundance relative to siControl. GAPDH was used as endogenous control. Data are mean ± s.e.m. For GM18453 (siControl n = 5, siPLA2G15_1 n = 5 and siPLA2G15_2 n = 6) and for GM03123 (siControl n = 6, siPLA2G15_1 n = 6 and siPLA2G15_2 n = 5). ****p < 0.0001, by one-way ANOVA Fisher’s LSD test.
