Fig. 3
Hypoxia Leads to Changes in H-EVs and N-EVs miRNA Profiles and miR-612 mediates the pro-angiogenic effects of OM-MSC-EVs on HBMECs. A Differential expression level of exosomal miRNAs between H-EVs and N-EVs. MiR-612 is one of the exosomal miRNAs with markably greater abundance in H-EVs compared to N-EVs. B Volcano plot of differentially expressed miRNAs between H-EVs and N-EVs. Red, significantly upregulated miRNAs in H-EVs; green, significantly downregulated miRNAs in N-EVs; gray, no significant difference. Fold change > 2 and P < 0.05 were considered significant. The sequencing samples of H-EVs and N-EVs were isolated from six sample. C KEGG pathway analysis of the target genes of five significantly upregulated miRNAs in H-EVs. Top 20 enriched pathways are indicated. D Expression level of miR-612 in H-OM-MSCs and N-OM-MSCs, (n = 9), ***P < 0.001. E MiR-612 abundance in EVs secreted by H-EVs and N-EVs, (n = 9), ***P < 0.001. F After 3 h of H-EVs or N-EVs treatment, the levels of miR-612 in HBMECs were measured by qPCR analysis. Data were normalized to levels of U6 (cellular) or total protein of EVs, (n = 9), *P < 0.05, ***P < 0.001. G Cell viability at 6 h and 12 h was examined in HBMECs that were treated with EVs obtained from OM-MSCs that were pretreated with scramble (OM-MSC-EVsNC)or with EVs isolated from OM-MSCs that were pretreated with anti-miR-612 (OM-MSC-EVsanti-miR-612), (n = 5),***P < 0.001. H The migration of HBMECs stimulated by OM-MSC-EVsNC and OM-MSC-EVsanti-miR-612 was detected by the transwell assay, (n = 9), ***P < 0.001, Scale bar: 100 μm. I Representative images of the tube formation assay in HBMECs treated with OM-MSC-EVsNC and OM-MSC-EVsanti-miR-612,Scale bar: 200 μm. J Quantitative analyses of the total junctions, meshes and nodes in (I), (n = 9), *P < 0.05, ***P < 0.001