Fig. 2
MSCpos-EVs modulate the phenotype and angiogenic ability of endothelial cells. A) Quantification of the number of intersections and network perimeters formed by HUVECs on the Matrigel layer after MSCpos- and MSCneg-EV treatment. Untreated cells were used as a control. (n = 5 per group). B) Flow cytometric analysis of the activated HUVEC phenotype (CD31+/CD34+/CXCR4+) after MSCpos- and MSCneg-EV treatment. Untreated cells (NT) were used as a control (n = 5). C) Tumor growth curves and CD31 IHC staining of 104 lung cancer (A549) cells coinjected with HUVECs pretreated with MSCpos- and MSCneg-EVs in immunodeficient mice (n = 7 for each group). D) Analysis of the relative expression levels of miR-126, pre-miR-126, SPRED1 and VEGF in HUVECs treated with MSCpos- and MSCneg-EVs. Untreated cells (NT) were used as a control (n = 5). E) Tube formation assay of miR-126-overexpressing HUVECs compared to control SCR-expressing HUVECs on Matrigel (n = 5). F) The graphs show the percentages of the CD31+/CD34+/CXCR4+ population among miR-126-overexpressing cells (n = 5) and (G) the in vivo growth curves of 105 A549 cells coinjected with the same HUVECs (n = 5 mice per group). H) Cells pretransfected with LNA 126 were incubated with MSCpos-EVs, and capillary-like structure formation was analyzed (n = 4). I) The panel shows a reduction in the CD31+/CD34+/CXCR4+ HUVEC-LNA126 population after treatment with MSCpos-EVs compared to the corresponding control treatment (n = 4). L) Vessel network formation and M) activation of endothelial cells after treatment with MSCneg-EVs and MSCneg-EVs enriched with mimic-126 (n = 5). *p < 0.05 versus controls. The data are expressed as the mean ± S.E.M. values