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. 2021 Feb 2;47(4):35. doi: 10.3892/ijmm.2021.4868

Figure 1.

Figure 1

Persistent exposure of HUVECs to HG increases miR-6832-5p, miR-6842-5p and miR-8056 expression by downregulating MIR181A2HG. (A) MTS assay detected the viability of HUVECs treated with different doses of D-glucose for the indicated times. **P<0.01 and ***P<0.001 vs. day 0. (B) Heatmap representation of differential expression of lncRNAs between HUVECs treated with 5.5 mM (L5.5) and 30 mM D-glucose (H30) for 24 h. (C) RT-qPCR was used to detect the relative expression of MIR181A2HG in HUVECs cultured in medium containing 30 mM D-glucose for the indicated times. *P<0.05 and ***P<0.001 vs. day 0. (D) FISH was used to detect the location and expression of MIR181A2HG in HUVECs exposed to 5.5 or 30 mM D-glucose for the indicated times. (E) Predicted binding sites of the miRNAs on MIR181A2HG. (F) Luciferase reporter assay was used to verify the direct binding between MIR181A2HG and miRNAs. *P<0.05 and **P<0.01 vs. miR-Ctl. (G) RT-qPCR was used to detect the relative expression of the indicated miRNAs in HUVECs cultured in medium containing 30 mM D-glucose for 96 h. *P<0.05 and **P<0.01 vs. Ctl. (H) RNA pulldown assay was performed to detect the direct binding between MIR181A2HG and the 3 miRNAs. **P<0.01 vs. Ctl. (I and J) RT-qPCR was used to detect the relative expression of (I) MIR181A2HG and (J) miRNAs in HASMCs cultured in medium containing 5.5 or 30 mM D-glucose. (K and L) RT-qPCR and western blot analysis detected the relative expression of AKT2 mRNA and protein in HASMCs transfected with miR-181-3p mimics, miR-181-5p mimics or their corresponding control. HUVECs, human umbilical vein endothelial cells; HASMCs, human aortic smooth muscle cells.