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. 2022 Nov 1;11(21):3457. doi: 10.3390/cells11213457

Figure 5.

Figure 5

miR-140-3p and miR-574-3p downregulate VEGF expression. (A) VEGF expression in the umbilical vein tissue of normal (n = 6) and GDM (n = 7) pregnant women was measured by Western blotting. (B) Linear regression analysis of miR-140-3p or miR-574-3p expression and VEGF in the umbilical vein tissue in pregnant women (n = 13). (C) VEGF expression of MUVECs after treatment with miRNA AG NC, miR-140-3p AG, miR-574-3p AG, miR-140-3p + miR-574-3p AG, miRNA Anta-AG NC, miR-140-3p Anta-AG, miR-574-3p Anta-AG, and miR-140-3p + miR-574-3p Anta-AG was measured by Western blotting (n = 3). (D) TargetScan and a protein–protein interaction (PPI) network were applied to identify the targets of miR-140-3p and their interaction. (E) Co-transfection of wild-type or mutant seed regions of VEGF 3′UTR were constructed with miRNA AG NC or miR-140-3p AG in MUVECs. A luciferase assay was applied to detect the luciferase activity (n = 3). (F) Protein levels of VEGF and hnRNP L after transfection with miRNA AG NC+vector and miR-574-3p AG+vector, miRNA AG NC+hnRNP L overexpression (OE), and miR-574-3p AG+hnRNPL OE were measured by Western blotting (n = 3). ** p < 0.01. ns: no significance.