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. 2022 Sep 12;13:984302. doi: 10.3389/fimmu.2022.984302

Figure 4.

Figure 4

miR-146aKO B-cells have upregulated interferon pathway genes expression and Traf6 is a target for miR-146a dependent activation. (A) Snapshot of heatmap from the RNA-Seq data showing differentially expressed IFN targeted genes. miR-146a molecular target, Traf6, is highlighted in yellow. (B) GSEA of genes differentially expressed between WT and miR-146aKO activated B-cells from LPS stimulation at 96 hours shows enrichment in Interferon Modules within SLE Disease. Normalized Enrichment Score: -1.94; FDR q value: 0.003. (C) Metascape enrichment analysis of differentially expressed genes between WT and miR-146aKO B-cells with LPS and (D) CpG stimulation show highest enrichment for genes within the Reactome Cytokine Signaling in Immune System and Interferon Signaling Pathways. (E) RT-qPCR validation for Traf6 expression with LPS and (F) CpG stimulation. One-way ANOVA with Tukey’s multiple-comparisons test; *P < 0.05, **P < 0.01, ***P < 0.001. (G) RT-qPCR of downstream Traf6 genes in LPS and CpG-stimulated B-cells. t test; *P < 0.05, **P < 0.01, ***P < 0.001. Data represent mean ± SEM. Graphs combine three independent experiments (n = 3-4 mice/group/exp).