Extended Data Fig. 5: Multiple TREM2 and PLCG2 KO iMG clones display shared lipid accumulation defects upon myelin challenge.
a, Lipidomic analysis of free cholesterol (left), cholesteryl ester (middle), and ceramide (right) species in second, independent TREM2 and PLCG2 KO iMG clones treated with vehicle or myelin (25 µg/ml) for 48 hours. Mean lipid abundance value for each species is normalized to cell number and presented on a log2 transformed scale. n = 4 biologically independent experiments, except PLCG2 KO conditions for ceramide species, n = 3 biologically independent experiments. b, Lipidomic analysis of hexosylceramide (left) and diacylglycerol (right) species in indicated iMG genotypes and clones treated with vehicle or myelin (25 µg/ml) for 48 hours. Mean lipid abundance value for each species is normalized to cell number and presented on a log2 transformed scale. n = 3 biologically independent experiments, except WT and TREM2 KO clone 2 conditions, n = 4 biologically independent experiments. c, Relative TREM2 mRNA expression levels in indicated iMG populations. n = 4 biologically independent experiments. d, Flow cytometry histogram shows TREM2 surface expression in TREM2(R47H) iMG. n = 3 biologically independent experiments, with similar results. e, Relative fraction of cells with pHrodo signal remaining 96 hours after treatment with pHrodo-myelin in the indicated iMG populations, normalized to WT as 1. TREM2(R47H), n = 2 biologically independent experiments; TREM2 HET, n = 3 biologically independent experiments; WT, TREM2 KO, n = 4 biologically independent experiments. f, Relative PLCG2 mRNA expression levels in indicated iMG populations. n = 4 biologically independent experiments. All data are mean ± s.e.m. For (a-b), two-way ANOVA, genotype effect between treatment populations, post-hoc Tukey test for multiple comparisons. All statistical tests were performed on log2 transformed abundance values. OE, transgene expression.
