Fig. 4
CRC-secreted miR-25-3p silences KLF2 and KLF4 in HUVECs. a Western blot analysis of KLF2, VEGFR2, AKT, p-AKT, ERK, p-ERK expression in HUVECs incubated with NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + Annexin V, SW480/miR-25-3p exosomes + miR-25-3p inhibitor and SW480/miR-25-3p exosomes + KLF2 groups. b Western blot analysis of KLF4, ZO-1, occludin, Claudin5 expression in HUVECs incubated with NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + Annexin V, SW480/miR-25-3p exosomes + miR-25-3p inhibitor and SW480/miR-25-3p exosomes + KLF4 groups. c Immunofluorescence staining analysis of ZO-1, occludin, Claudin5 expression in HUVECs incubated with NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + Annexin V, SW480/miR-25-3p exosomes + miR-25-3p inhibitor and SW480/miR-25-3p exosomes + KLF4 groups. Scale bars represents 10 µm. d Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + Annexin V, SW480/miR-25-3p exosomes + miR-25-3p inhibitor and SW480/miR-25-3p exosomes + KLF4 treatments on tube formation ability of HUVECs by tube formation assay. Scale bar represents 100 µm. Mean ± SEM are provided (n = 3). e Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + Annexin V, SW480/miR-25-3p exosomes + miR-25-3p inhibitor and SW480/miR-25-3p exosomes + KLF2 treatments on vascular outgrowth of rat aortic rings. Scale bar represents 200 µm. Mean ± SEM are provided (n = 3). f Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + Annexin V, SW480/miR-25-3p exosomes + miR-25-3p inhibitor and SW480/miR-25-3p exosomes + KLF4 treatments on permeability of HUVEC monolayers by in vitro permeability assay. Mean ± SEM are provided (n = 3). **P < 0.01, ***P < 0.001 according to two-tailed Student’s t test
