Fig. 6
Exosomal miR-467b-3p levels are associated with macrophage polarization. A Hierarchical cluster heatmap of the differential miRNAs in UTX−/−-ECs compared to UTXf/f-ECs. Each cell represents a differentially expressed miRNA. The color scale from green (low) to red (high) indicates the expression levels of DEPs. B Volcano plot of differentially expressed miRNA (P value < 0.05 and log2 Fold-Change > 1) UTX−/−-ECs compared with UTXf/f-ECs. C The Top10 miRNAs family of differentially expressed miRNAs. D The expression levels top10 up-regulated candidate miRNAs. E The miRNAs’ expression level in UTX−/−-Exos verified by qRT-PCR. *P < 0.05, #P < 0.01 vs. UTXf/f-Exos. F The enrichment of miRNAs in UTX−/−-Exos verified by qRT-PCR. nsP>0.05, *P<0.05, **P<0.01 vs. UTX−/−-ECs. G–L qRT-PCR analysis of the expression of M1 markers (iNOS2, TNF-ɑ, and CD86) and M2 markers (Arginase-1, IL-10, and CD206) of LPS-stimulated macrophages after being treated with the mimics and inhibitors of the miRNAs (miR-467b-3p, miR-7224-3p, miR-346-5p, and miR-466l-3p). M The iNOS and Arginase-1 expression of LPS-stimulated macrophages when exposed to the mimic and inhibitor of miR-467b-3p compared to the control mimic/inhibitor. N, O Statistical analysis of Arginase-1 and iNOS expression in M. P Immunofluorescent analysis of LPS-stimulated macrophage (green) phenotypes (red) when exposed to the mimic and inhibitor of miR-467b-3p compared with control mimic and inhibitor. Scale bar, 20 μm. Q, R Statistical analysis of mean fluorescence intensity of CD86 and CD206 in P. nsP > 0.05, *P < 0.05, **P < 0.01 vs. Control mimic/inhibitor. n = 6/per group