Figure 3.

miR-145 Increases Endothelial Angiogenic Functions

(A) Representative images of vascular sprouting from aortic rings isolated from P20 mice. Tissue explants were treated with a non-targeting negative control (left) or the miR-145 mimic (right). (B) Quantitative analysis of the sprouting area in day 4 explants showed that the miR-145 mimic significantly increased sprouting ability ex vivo compared with rings treated with the negative control (n = 6–9 per group). (C–F) Human retinal microvascular endothelial cells (HRMECs) were treated with the miR-145 mimic, inhibitor, or negative control and analyzed for endothelial functions. Representative images (C) show tubular formation (top) and results of a wound-healing migration assay (bottom; white dashed lines indicate the initial boundaries of the scratches, white solid lines show the leading edges at 24 h after treatment, and the yellow arrow indicates the direction of cell migration). Quantitative analyses show that HRMECs treated with the miR-145 mimic displayed higher levels of proliferative activity in an MTT assay (D), mesh areas in a tube-formation assay (E), and cell coverage areas during migration (F), compared with cells treated with the negative control. Inhibition of miR-145 in HRMECs reveals significantly decreased levels of tube formation (E) and migration (F). (D) n = 3 per condition; (E and F) n = 6 per group. Scale bars represent 1 mm (A) and 100 μm (C). Neg Ctrl, negative control RNA oligomers. Data are presented as means ± SEM. *p ≤ 0.05; **p ≤ 0.005; ***p < 0.001.