Figure 2.

Exosomal EPHA2 promotes endothelial cell angiogenesis. A Expression of EPHA2 in exosomes and cell lysates was analyzed by using Western blotting; β-actin was used as the loading control. B Western blot analysis showed that EPHA2 expression was silenced in high metastatic potential cells and exosomes after infection with lentivirus expressing EPHA2-specific shRNAs; β-actin was used as the loading control. C, D EPHA2-silenced HM-exos failed to promote the tube formation of endothelial cells and the microvascular outgrowth of rat arterial rings. E Matrigel plug assay showed that exosomal EPHA2 promotes angiogenesis in vivo. For the in vivo matrix gel plug assay, Matrigel plugs with or without VEGF were used as positive or negative controls. HE and CD31 staining were carried out on the dissected Matrigel plugs. F, G EPHA2-silenced HM-exos failed to promote the migration of endothelial cells as measured by Transwell and Wound healing assays. H Western blot analysis showed the expression of EPHA2 in HEK-293T cells and exosomes in cells that were transfected with control or EPHA2 expression vectors. β-actin was used as the loading control. I, J Exosomes from HEK-293T cells expressing EPHA2 significantly promote the tube-forming capacity of endothelial cells compared with control exosomes. Data are expressed as mean ± SD, and all experiments were repeated at least three times. *P < 0.05, **P < 0.01, ***P < 0.001 and ns P > 0.05 indicate no statistical significance. Scale bar: 200 μm.