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. 2020 Apr 21;22(1):483–493. doi: 10.3892/mmr.2020.11085

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Copyright: © Ma et al.

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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Figure 2. — miR-93 enhances endothelial cell migration, and suppresses Lats2 expression and Yap phosphorylation in endothelial cells. (A) Representative images of scratch-wound assay before and 12 h after scratch formation in HUVECs (magnification, ×200). The line indicates the width of the gap. Transfection with miR-93 mimic promoted HUVEC migration into the wound area. n=4 replicates per group. (B) Predicted binding sites between miR-93 and Lats2 3′UTR. miR-93 mimics decreased luciferase activity. n=4 replicates per group. (C) Representative bands of Lats2, p-Yap and Yap in HUVECs. Expression of (D) Lats2 was suppressed in miR-93 mimic-transfected HUVECs. Transfection with miR-93 mimic (E) decreased Yap phosphorylation and (F) increased Yap expression in the cytosol after HUVECs H/R. n=4 replicates per group. HCMECs were transfected with miR-93 mimics or scr-miR mimics for 48 h. (G) Reverse transcription-quantitative PCR showed the upregulation of miR-93 in HCMECs by miR-93 mimics transfection. n=4 replicates per group. (H) Transfection of HCMECs with miR-93 mimic increased cell viability following H/R injury. n=4 replicates per group. (I) Representative images of scratch-wound assay at 0 and 12 h after scratch formation in HCMECs (magnification, ×100). The line indicates the width of the gap. Transfection with miR-93 mimic promoted HCMEC migration into the wound area. n=4 replicates per group. (J) Representative bands of Lats2 and p-Yap in HCMECs. Western blotting results identified the attenuation of (K) Lats2 and (L) p-Yap expression levels by miR-93 transfection after H/R in HCMECs. Data are presented as the mean ± SEM. Each experiment was repeated three times. *P<0.05 vs. the indicated group. Lats2, large tumor suppressor 2; Yap, Yes-associated protein; HUVECs, Human umbilical vein endothelial cells; HCMECs, human cardiac microvascular endothelial cells; scr, scrambled; miR, microRNA; H/R, hypoxia/reoxygenation; p, phosphorylated; 3′UTR, 3′ untranslated region; WT, wild-type; MUT, mutant.

Figure 2. — miR-93 enhances endothelial cell migration, and suppresses Lats2 expression and Yap phosphorylation in endothelial cells. (A) Representative images of scratch-wound assay before and 12 h after scratch formation in HUVECs (magnification, ×200). The line indicates the width of the gap. Transfection with miR-93 mimic promoted HUVEC migration into the wound area. n=4 replicates per group. (B) Predicted binding sites between miR-93 and Lats2 3′UTR. miR-93 mimics decreased luciferase activity. n=4 replicates per group. (C) Representative bands of Lats2, p-Yap and Yap in HUVECs. Expression of (D) Lats2 was suppressed in miR-93 mimic-transfected HUVECs. Transfection with miR-93 mimic (E) decreased Yap phosphorylation and (F) increased Yap expression in the cytosol after HUVECs H/R. n=4 replicates per group. HCMECs were transfected with miR-93 mimics or scr-miR mimics for 48 h. (G) Reverse transcription-quantitative PCR showed the upregulation of miR-93 in HCMECs by miR-93 mimics transfection. n=4 replicates per group. (H) Transfection of HCMECs with miR-93 mimic increased cell viability following H/R injury. n=4 replicates per group. (I) Representative images of scratch-wound assay at 0 and 12 h after scratch formation in HCMECs (magnification, ×100). The line indicates the width of the gap. Transfection with miR-93 mimic promoted HCMEC migration into the wound area. n=4 replicates per group. (J) Representative bands of Lats2 and p-Yap in HCMECs. Western blotting results identified the attenuation of (K) Lats2 and (L) p-Yap expression levels by miR-93 transfection after H/R in HCMECs. Data are presented as the mean ± SEM. Each experiment was repeated three times. *P<0.05 vs. the indicated group. Lats2, large tumor suppressor 2; Yap, Yes-associated protein; HUVECs, Human umbilical vein endothelial cells; HCMECs, human cardiac microvascular endothelial cells; scr, scrambled; miR, microRNA; H/R, hypoxia/reoxygenation; p, phosphorylated; 3′UTR, 3′ untranslated region; WT, wild-type; MUT, mutant.