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. 2021 Jul 16;9(7):830. doi: 10.3390/biomedicines9070830

Figure 3.

Figure 3

MiR overexpression in endothelial tissue. (A) Transfection of HUVEC cells with miR mimic: Since miR-181/-429 were upregulated in COPD lung, a mimic was used to test function. The mimic successfully upregulated the miR in HUVECs 3 days after transfection (n = 3 in both groups), *** = p < 0.001. (B) Number of nodes formed by HUVECs imbedded in Matrigel after transfection of miR-429 mimic. The figure shows the number of nodes present 3, 6, 18 and 24 h after HUVECs were seeded on Matrigel. The numbers represent the mean from 3 separate experiments. An ANOVA test was used to look for significance between groups. * = p < 0.05 (ANOVA); *** = p <0.001. Tukey’s test was used to compare groups, which demonstrated that number of nodes was significantly reduced in the mimic group in comparison to the negative siRNA group at 3 h (p < 0.001), suggesting that miR-429 reduces angiogenesis. (C) Total mesh area formed by HUVECs embedded in Matrigel after transfection of miR-181b-3p mimic: the figure shows the total mesh area present 6, 12, 18 and 24 h after HUVECs were seeded on Matrigel. The numbers represent the mean from 3 separate experiments. An ANOVA test was used to look for significance between groups. ** = p < 0.01 (ANOVA); *** = p <0.001. Tukey’s test was used to compare groups, which demonstrated that total mesh area was significantly reduced in the mimic group in comparison to the negative siRNA group at 6 (p < 0.01 and 12 h (p = 0.042), suggesting that miR-181b-3p reduces angiogenesis. Representative images from these experiments are shown; the picture on the left shows HUVECs at 12 h transfected with the miR mimic, and on the right with the negative siRNA. Scale bars represent 800 μm. (D) Number of endothelial sprouts from spheroids after transfection of a miR-181b-3p mimic: the figure shows the total number of endothelial sprouts for each condition. The numbers represent the mean from 3 separate experiments. An ANOVA test was used to look for significance between groups. *** = p <0.001 (ANOVA). Tukey’s test was used to compare groups, which demonstrated that number of sprouts was significantly reduced in the mimic group in comparison to the negative siRNA group. Representative images from these experiments are shown; the picture on the left shows HUVECs transfected with the miR mimic, and on the right with the negative siRNA. Scale bars represent 200 μm.