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. 2021 Jul 13;9:92–104. doi: 10.1016/j.bioactmat.2021.07.007

Fig. 3.

Fig. 3

Effect of NGO on angiogenesis in vitro. (a) HUVECs were incubated in endothelial culture medium (basal endothelial culture medium supplemented with 10% FBS) with NGO at a concentration gradient for 6, 24 and 48 h, and a CCK-8 assay was performed to determine cell viability. (b) Schematic of tip-stalk cell selection in angiogenesis. (c) Wound healing assay in HUVECs cultured in endothelial culture medium with 0, 1, 5, 20 or 50 μg/mL NGO for 16 h. The wound area (%) was measured using ImageJ. (d) Tube formation by HUVECs cultured in endothelial culture medium with 0, 1, 5, 20 or 50 μg/mL NGO for 24 h. The quantified master segment length (μm) and numbers of nodes and meshes are presented in a histogram. (e) The mRNA expression of tip cell-associated genes in HUVECs cultured in endothelial culture medium with 0, 1, 5, 20 or 50 μg/mL NGO for 24 h. (f) The mRNA expression of stalk cell-associated genes in HUVECs treated with 0, 1, 5, 20 or 50 μg/mL NGO for 24 h. (g) Immunoblot detection of KDR, DLL4, and CD34 in HUVECs treated with NGO for 24 h. Data represent the mean ± SD (n = 3). *p < 0.05, **p < 0.005, ***p < 0.0001 compared with the control group.