Fig. 7
Sulfatide deficiency-induced astrogliosis and upregulated ApoE were not secondary to microgliosis. CST+/+ and CST−/− mice were fed PLX3397-containing or control chow-like diet from 1 to 3 mo of age. (A) Representative immunofluorescence images from brain sections using antibodies against Iba-1 (green) and GFAP (red); nuclei were stained with DAPI; See Fig. S10A for higher magnification and merged images. Scale bar: 500 μm. (B-D) Brain mRNA from four groups (CST+/+, CST+/++PLX3397, CST−/−, and CST−/− + PLX3397 mice) was accessed using NanoString neuroinflammation panel. (B) Volcano plot displaying -log10 p-value and log2 fold change for each gene to show the CST KO effect (middle) and drug-effect within CST+/+ (left) and CST−/− (right) brain. See Fig. S10B-D for PCA and Venn diagrams showing the number of specific and shared DEGs between treatments. (C) NanoString pathway score analysis for astrocyte function, microglia function, innate immune response, and inflammatory signaling for each of the four groups. Two-way ANOVA with Bonferroni post-hoc test for multiple comparisons, n = 3. (D) Fold changes of several typical microgliosis or astrogliosis related genes in CST+/+ and CST−/− brain with or without PLX3397 treatment to display the drug effects on microglia and astrocytes. Heteroscedastic Welch’s t-Test, n = 3. (E) Western blot analysis from cerebrum homogenates using antibodies against Iba-1, GFAP, ApoE and β-actin. Two-tailed unpaired t-Test, n = 3. #p < 0.1, *p < 0.05, **p < 0.01, ***p < 0.001. Data represent the mean ± S.E.M