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. 2021 Apr 20;9:619444. doi: 10.3389/fcell.2021.619444

FIGURE 3.

FIGURE 3

miR-185 directly targets CXCL12 in ECs. (A) The predicted miR-185 target site in the 3′ UTR of CXCL12. (B) HEK293 cells were co-transfected with miR-185 mimic or inhibitor together wildtype (wt)- or mutant (mut)-CXCL12 3′ UTR construct. Twenty-four hours later, cells were harvested for the determination of luciferase activities. **P < 0.01 vs. NC or NIC group. (C) Western blot analysis of CXCL12 in ECs treated with different media. *P < 0.05 vs. Basal-CM. (D) Western blot analysis of CXCL12 in ECs transfected with miR-185 mimics and inhibitor for 48 h. (E) Proliferation of ECs transfected with miR-185 mimics and inhibitor was assessed by a BrdU assay. (F) Migration of ECs transfected with miR-185 mimics and inhibitor was assessed by a scratch wound assay. Quantification of migration is expressed as a relative rate of scratch wound healing. (G) Angiogenesis of ECs transfected with miR-185 mimics and inhibitor was assessed by a tube formation assay. Quantitative analysis of the tube lengths is shown. *P < 0.05, **P < 0.01 vs. NC or NIC group. (H–K). ECs transfected with miR-185 inhibitor were treated with AMD3100 (5 μM) for 24 h. Western blot analysis of CXCR4 (H), proliferation (I), migration (J), and angiogenesis (K) of ECs transfected with miR-185 inhibitor and treated with AMD3100 were assessed. *P < 0.05, **P < 0.01 vs. Blank, #P < 0.05, ###P < 0.001 vs. Inhibitor.