EZH2 Mediates miR-146a-5p/HIF-1α to Alleviate Inflammation and Glycolysis after Acute Spinal Cord Injury

Figure 4

EZH2 indirectly regulated the HIF-1α expression by mediating miR-146a-5p. (a) Western blot detected the expression of H3K27me3 in the Control group and LPS group. vs. Control group. (b) qRT-PCR detected miR-146a-5p expression in the Control group and LPS group. vs. Control group. (c) ChIP was used to analyze the enrichment of H3K27me3 in the miR-146a-5p promoter region. vs. Control group. (d) starBase predicted the binding sites of HIF-1α and miR-146a-5p, and dual-luciferase report assay verified the binding of HIF-1α and miR-146a-5p. vs. WT-HIF-1α group. (e) Western blot detected the expression of HIF-1α in the NC-EZH2 group, NC-miR-146a-5p group, miR-146a-5p inhibitor group, EZH2 inhibitor+NC-miR-146a-5p group, and EZH2 inhibitor+miR-146a-5p inhibitor group. vs. NC-EZH2 group; ^# vs. NC-EZH2+miR-146a-5p inhibitor group; ^& vs. NC-miR-146a-5p+EZH2 inhibitor. (f) Western blot detected inflammatory factors IL-6, TNF-α, and IL-17 and glycolysis-related gene PKM2, GLUT1, and HK2 expressions. (g) Inflammatory factors IL-6, TNF-α, and IL-17 and glycolysis-related gene PKM2, GLUT1, and HK2 expressions. vs. NC-EZH2 group; ^# vs. NC-EZH2+miR-146a-5p inhibitor group; ^& vs. NC-miR-146a-5p+EZH2 inhibitor.

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