Figure 5.

Exosomal miR-155-5p and miR-221-5p Promote Angiogenesis in an E2F2-Dependent Manner

(A) Protein expression analysis of E2F2 in MAECs 72 h after transfection with miR-155-5p mimics, miR-221-5p mimics, or the negative control (mir-NC) and miR-155-5p inhibitor, miR-221-5p inhibitor, or the negative control (Inh-NC). (B) Protein levels of E2F2 after 72-h incubation of MAECs with exosome-free medium or M2-exo. (C) E2F2 protein levels of MAECs after incubation with M2-exo, miR-155-5p-inhibitor, and/or miR-221-5p-inhibitor. (D) In the 3′ UTR region of E2F2, the predicted binding region and mutation of mir-155-5p and mir-221-5p are shown. (E) miR-221-5p-inhibitor or miR-155-5p-inhibitor and plasmid containing the E2F2 3′ UTR region were co-transfected into MAECs, and their relative luciferase activities were detected. (F) miR-155-5p or miR-221-5p and plasmid containing the E2F2 3′ UTR region were co-transfected into MAECs, and their relative luciferase activities were detected. (G) Plasmids containing the wild-type E2F2 3′ UTR or its mutation at the predicted miR-155-5p and/or miR-221-5p target sequences were transfected into MAECs, along with M2 macrophage-derived exosomes, and the relative luciferase activities were detected. (H) Representative micrographs of the transwell assay (original magnification, ×100). The numbers of cells were calculated per high-power field from three independent experiments. (I) Representative micrographs of the tube formation assay (original magnification, ×200). The numbers of branch points were calculated by ImageJ. (J) Representative micrographs of the 24-h average distance of the wound-healing assay. (K–M) Data on transwell, wound-healing, and tube formation assays. ∗p < 0.05, ∗∗∗p < 0.001.