Fig. 5

Asrij promotes inflammatory disease-associated microglia (DAM) activation state. A Representative renderings of the microglial skeleton using the ‘filament’ module of Imaris. Graphs show quantification of branch number per microglia and total branch length (n = 3 for WT and KO, n = 6 for AD and AD/KO, all males). B Heatmap of significantly downregulated (P < 0.01, log₂FC < − 1) genes belonging to DAM signature. C Representative confocal images showing DAM marker, CLEC7A (magenta) on IBA1⁺ microglia (red) associated with ThioS⁺ plaques (green). Graphs show quantification of percent CLEC7A⁺ area normalized to IBA1⁺ area and percent CLEC7A⁺ area per plaque averaged over 15 plaques per section with 3 sections per mouse (n = 6, all males). D Representative confocal images showing homeostatic marker, TMEM119 (red) on IBA1⁺ microglia (magenta) associated with 6E10⁺ plaques (green). Graphs show quantification of percent TMEM119⁺ area normalized to IBA1⁺ area and percent TMEM119⁺ area per plaque averaged over 15 plaques per section with 3 sections per mouse (n = 6, all males). E, F Heatmaps of significantly downregulated (P < 0.01, log₂FC < − 1) pro-inflammatory genes and IRM signature genes. G, H, I Graphs show quantification of IL-6, TNF-α and IL-1β levels (pg/mL) in the hippocampus by ELISA (n = 6, all males). J Bar graph shows the top 10 significantly downregulated MSigDB hallmark pathways (P < 0.05). K Immunoblot analysis of phospho-STAT3 (tyrosine 705) and STAT3 levels in CD11b⁺ microglia. GAPDH is loading control. Graph shows quantification of fold change in p-STAT3 normalized to total STAT3 (n = 3, all females). L Immunoblot analysis of phospho-NF-κB (serine 563) and NF-κB levels in CD11b⁺ microglia. GAPDH is loading control. Graph shows quantification of fold change in p-NF-κB normalized to total NF-κB (n = 3, all females). Statistical significance between experimental groups was calculated by one-way ANOVA with Tukey’s post-hoc test (A, G, H and K, L), unpaired two-tailed Student’s t-test (C–D and I), and Fischer exact test (J). Error bars denote mean ± SEM. ns: non-significant, *P < 0.05, **P < 0.01, and ***P < 0.001