Extended Data Fig. 10: Cholesterol regulates astrocyte reactivity extended data.

a) STRING analysis of all proteins significantly upregulated >1.5 fold in Kolf2.1J and BIONi037 reactive astrocytes (dataset from proteomics in Fig. 5). Zoom in shows cluster of proteins related to cholesterol metabolism. b) Normalized Il-6 secretion in iAstrocytes (ApoE3), pre-treated with vehicle or avasimibe (0.5μM) for one hour and then treated for 24 hours with vehicle, cholesterol (50μM) or cholesterol + avasimibe. N=6, four or two independent experiments from two ApoE3 lines, Mean *P<0.05, ***P<0.001 one sample t-test for Chol and Ava + Chol vs 1. *P<0.05 t-test for Chol vs Ava + Chol with Benjamini-Hochberg correction on the three p-values. c) Relative mRNA expression of indicated genes after cholesterol (10μM) + avasimibe (0.5μM) treatment vs control, quantified by qPCR. N=5, two or three independent experiments from two ApoE3 lines, Mean + sem *P<0.05 one sample t-test with Benjamini-Hochberg correction on the p-values. d) Volcano plots represent log2fold change of individual lipid species in cholesterol treated vs control iAstrocytes in three independent replicate lipidomics experiments (BIONi037-A). e) Summary data of changes in all detected lipid classes in cholesterol vs control treated iAstrocytes (BIONi037-A). N=3 Median + IQR. Paired t-test or paired Mann Whitney U test with Benjamini Hochberg correction. f) Bar graphs of lipid classes in cholesterol vs control treated iAstrocytes from BIONi037-A background (% of total). All experiments presented in E are included here. Mean + sd. g-h) Fold change of phospholipid (g) and triacylglyceride (h) species with indicated number of double bonds in cholesterol treated vs control iAstrocytes (BIONi037-A). All experiments presented in E are included here. Mean. In all panels open circles or triangles indicate mean per experiment, while solid dots represent all independent wells.