Fig. 2.

AZIN2-sv induces endothelial apoptosis and inhibits vessel formation.

(A) Flow cytometric analysis of HUVEC apoptosis was performed forty-eight hours after transfection with shAZIN2-sv. H₂O₂ (400 μM) was used to induce apoptosis of endothelial cells. (B) Quantification of apoptotic endothelial cells. ^⁎p < .05 vs. control. ^#p < .05 vs. H₂O₂; n = 6 per group. (C) HUVEC proliferation was analysed by flow cytometry. (D) Quantification of Median Fluorescence Intensity (MFI). ^⁎p < .05 vs. vector; n = 6 per group. (E) Representative images of vascular sprouting. Cells were treated with or without VEGF (10 ng/mL). Forty-eight hours after transfection with control vector or AZIN2-sv, HUVEC spheroids were allowed to undergo sprouting in a 3-dimensional (3D) matrix for 24 h (bars, 500 μm). (F) Quantification of the cumulative length of sprouts per spheroid. ^⁎p < .05 vs. control. (G) HUVECs were seeded on Matrigel matrix and stimulated with TNF-α (10 ng/mL). Formation of tube-like structures was observed 24 h after TNF-α treatment. The average number of tubes formed per field was statistically analysed (bars, 500 μm). (H) Quantification of the average number of tubes. ^⁎p < .05 vs. TNF-a. ^#p < .05 vs. TNF-a + AZIN2-sv; n = 6 per group.