Fig. 4From: Regulation of human microglial gene expression and function via RNAase-H active antisense oligonucleotides in vivo in Alzheimer’s diseasePhenotypic and transcriptomic changes in human microglia upon ASO mediated APOE and TREM2 knockdown. A,B APOE (A) and TREM2 (B) expression in human microglia isolated from 6-month-old AppNL−G−F mice treated with 45ug APOE ASO-1 (A) or 90ug TREM2 ASO-171 (B) for 1 or 4 weeks. C ASO concentration in human microglia isolated from 6-month-old AppNL−G−F mice treated with APOE ASO-1 or TREM2 ASO-171 for 1 or 4 weeks. CNRQ = Calibrated Normalized Relative Quantities. Dots or bars represent mean ± SEM, n = 4/group. Statistical differences based on Two-way ANOVA test: *p < 0.05, **p < 0.01. D Representative images showing activated human microglia targeting X-34 positive amyloid-β fibrils 4 weeks post APOE and TREM2 knockdown. Arrow indicates an instance of activated human microglia around plaques. Scale bars = 100 µm (E) Quantification of mean fluorescence intensity for hP2RY12 and hCD9 at varying distances from the plaque assessed 4 weeks following ASOs treatment. F Quantification of hCD9 intensity at the plaque site (10 µm) normalized against signal intensity at the distal site from the plaque (74 – 81 µm) 4 weeks post ASOs treatment. Analysis was restricted to plaques surrounded by engrafted human microglia, determined by hP2RY12 signal (see Suppl Fig. 11A). Data represented as mean ± SD, n = 3/group. Statistical significance was evaluated with One-way ANOVA test (*p < 0.05, ns, not significant). G Gene set enrichment analysis (GSEA): Pre-ranked results of the microglial subtype gene sets (top 50 most significantly up-regulated genes of each microglial subtype [22]). The color represents the Normalized Enrichment Score (NES) of each gene set against the full list of genes ranked by the log-fold change when comparing the treatment group against the vehicle group. The asterisk indicates gene sets that are significantly enriched with FDR *p < 0.05. All analysis were performed on xenotransplanted microglia isolated from 6–7-month-old AppNL−G−F mice treated with APOE ASO-1 or TREM2 ASO-171 for 1 or 4 weeksBack to article page