0.5 MPa/NC-Exos Silence Wnt11 and Inhibit Angiogenesis in HUVECs
(A) Luciferase activities of 3′ UTR WNT11-luc constructs in HUVECs after incubation with 0 MPa/NC-exos, 0.5 MPa/NC-exos, 1.0 MPa/NC-exos, 0.5 MPa/NC-exos + Annexin V, and 0.5 MPa/NC-exos + miR-140-5p inhibitor. Mean ± SEM is provided (n = 3). ∗p < 0.05, ∗∗p < 0.01 for comparison with the 0.5-MPa/NC-exos group. (B) Western blotting analysis of Wnt11, β-catenin of nucleus/cytoplasm in HUVECs after incubation with 0.5 MPa/NC-exos, 0.5 MPa/NC-exos + Annexin V, 0.5 MPa/NC-exos + miR-140-5p inhibitor, and 0.5 MPa/NC-exos + Wnt11. (C) Immunofluorescence staining analysis of β-catenin and PKH67 lipid dye of HUVECs after incubation with 0.5 MPa/NC-exos, 0.5 MPa/NC-exos + Annexin V, 0.5 MPa/NC-exos + miR-140-5p inhibitor, and 0.5 MPa/NC-exos + Wnt11. β-catenin expression was decreased in the nucleus of the 0.5-MPa/NC-exos group. Bar, 10 μm. (D) ELISA detection of MMP-2 and MMP-7 levels in medium of HUVECs incubated with 0.5 MPa/NC-exos, 0.5 MPa/NC-exos + Annexin V, 0.5 MPa/NC-exos + miR-140-5p inhibitor, and 0.5 MPa/NC-exos + Wnt11. Mean ± SEM is provided (n = 3). ∗p < 0.05, ∗∗p < 0.01 for comparison with the 0.5-MPa/NC-exos group. (E–H) Migration (E and F), tube formation (E and G), and proliferation of HUVECs (E and H) after incubation with 0.5 MPa/NC-exos, 0.5 MPa/NC-exos + Annexin V, 0.5 MPa/NC-exos + miR-140-5p inhibitor, and 0.5 MPa/NC-exos + Wnt11. Bar, 100 μm. Mean ± SEM is provided (n = 3). ∗p < 0.05, ∗∗p < 0.01 for comparison with the 0.5-MPa/NC-exos group.