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. 2021 Oct 27;12:751667. doi: 10.3389/fphar.2021.751667

FIGURE 3.

FIGURE 3

Confirmation of the circHIPK3/miR-149-5p/FOXO1/VEGF functional module in vitro. (A–B) FOXO1/VEGF mRNA (A) and protein expression (B) were obviously down-/up-regulated by miR-149-5p mimics/inhibitor (n = 3; *p < 0.05, **p < 0.01). (C–D) Co-effects of circHIPK3 and miR-149-5p on FOXO1/VEGF expression in RA-FLS were evaluated. FOXO1/VEGF mRNA (C) and protein. (D) The expressions were significantly elevated/reduced by circHIPK3 overexpression/miR-149-5p mimics compared with those of each control group. Moreover, the significant downregulation of FOXO1/VEGF expression repressed by miR-149-5p mimics could be obviously rescued by circHIPK3 overexpression; in other words, the significant upregulation of FOXO1/VEGF expression induced by circHIPK3 overexpression could be significantly inhibited by miR-149-5p mimics (n = 3; *p < 0.05, **p < 0.01, ***p < 0.001). (E–F) The mRNA (E) and protein (F) expression of VEGF in RA-FLS were significantly elevated/reduced by FOXO1 overexpression/knockdown (n = 3; *p < 0.05, **p < 0.01). (G–H) FOXO1 overexpression significantly upregulated VEGF mRNA (G) and protein (H) expression, and the increment of VEGF expression induced by FOXO1 overexpression could be significantly suppressed by miR-149-5p mimics (n = 3; *p < 0.05). Results are expressed as the mean ± S.E.M. CTL = PBS control group. si-scrl = scramble siRNA. NC = negative control.