Fig. 3

MSCpos-EVs induce M2 polarization of macrophages. A) Intracellular expression levels of miR-320, pre-miR-320 and M2 phenotype markers in macrophages treated with MSCpos- and MSCneg-EVs, as measured by qPCR. Untreated cells were used as a control (n = 5). B) Flow cytometric analysis of CD163 and CD206 expression in macrophages treated with MSCpos- and MSCneg-EVs (n = 5). C) Absolute quantification (copies/µl) of the miR-320 content in PMN-EVs isolated from MSCpos and MSCneg individuals (n = 9). D) Relative expression levels of miR-320, pre-miR-320, and M2 mRNA (center), as measured by qPCR, and CD163 and CD206 levels, as measured by flow cytometric analysis (right) of macrophages treated with MSCpos- and MSCneg-PMN-EVs (n = 4). E) Analysis of relative miRNA and mRNA expression levels in macrophages transfected with LNA-320 and treated with MSCpos-EVs (n = 4). F) Analysis of miR-320, mRNA and CD163/CD206 expression levels in macrophages treated with MSCneg-PMN-EVs enriched with the miRNA-320 mimic (n = 5). Untreated (NT) macrophages were used as a control. *p < 0.05 versus controls. The data are expressed as the mean ± S.E.M. values