Fig. 4

Asrij deficiency upregulates metabolic genes and increases mitochondrial activity in AD microglia. A Schematic representing the experimental workflow of microglia isolation for RNA-seq analysis (n = 5 mice, all males). B Volcano plot depicting differentially expressed genes (DEGs). Points in red and blue are significantly (P < 0.01) upregulated (log₂FC > 1) and downregulated (log₂FC < − 1) genes, respectively. C Bar graph shows top 10 significantly enriched gene ontology terms and their normalized enrichment score (NES) in upregulated DEGs (FDR < 0.05). D Bar graph shows top 10 significantly enriched gene ontology terms and their normalized enrichment score (NES) in downregulated DEGs (FDR < 0.05). E Heatmap of significantly upregulated (P < 0.01, log₂FC > 1) genes involved in oxidative phosphorylation. F Representative histograms show flow cytometry analysis of Mitotracker Deep Red, TMRM, and MitoSOX Red staining in CD11b⁺ CD45^int microglia. Black dotted line represents fluorescence minus one (FMO) control. Graphs show quantification of fold change in median fluorescence intensity (MFI) of the indicated mitochondrial dyes (n = 5, all females). Statistical significance between experimental groups was calculated by two-sided Wald test (B), hypergeometric test (C-D) and one-way ANOVA with Tukey’s post-hoc test (F). Error bars denote mean ± SEM. ns- non-significant, *P < 0.05, **P < 0.01, and ***P < 0.001