Fig. 4
Methoxy-X04 (+) microglia from AppSAA KI/KI mice exhibit exacerbated gene expression and metabolic profiles. a (Left) Confocal images of cortex from 8-month-old AppSAA homozygous mice (KI/KI) stained for amyloid plaques and microglia markers (Iba1) post methoxy-X04 injection. Scale bars = 25 μm. (Right) a magnified view of the inset. b Quantification of methoxy-X04 (+) microglia proximal to plaques n=6. c-d Schematic of FACS experiment and gating strategy used to isolate pure populations of microglia that are negative (-) or positive (+) for methoxy-X04. e Fraction of methoxy-X04 (+) microglia purified from WT and AppSAA KI/KI mouse brains at 8 months of age. N = 10. f Volcano plot showing log2 fold change of gene expression between methoxy-X04 (+) and methoxy-X04 (-) samples. Genes expressed higher in methoxy (+) microglia have log2 fold changes > 0; colors same as in Fig. 3b. g Images of brain sections showing methoxy-X04 labeling and detection of Trem2 and Tmem119 transcripts by in situ hybridization. h Gene expression profiles from a representative subset of differentially expressed genes in methoxy-X04 (-) and (+) AppSAA KI/KI microglia when compared to WT. Intensities correspond to the log2 fold- change for each gene (row) per sample (column) as compared to the mean expression of the gene in the methoxy-X04 (-) WT group. i Gene set enrichment analysis from methoxy-X04 (-) (grey) or methoxy-X04 (+) (light green) AppSAA KI/KI vs methoxy-X04 (-) WT mice. Enrichment scores are calculated as the mean t-statistic of genes in the leading edge of the gene set. Filled bars indicate gene set enrichment results with FDR <= 10%