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. 2022 Nov 30;135(20):2467–2477. doi: 10.1097/CM9.0000000000002377

Figure 5.

Figure 5

TRAF6 is a direct target of miR-590-3p. (A) Schematic representation of the potential miR-590-3p binding sites in the 3′ UTR of TRAF6 as predicted by the online database. (B) Relative luciferase activity of the wild type (WT) and mutated (MUT) reporter constructs cotransfected with either the miR-590-3p mimic or scrambled oligonucleotides. (C) After the transfection of miR-590-3p inhibitors into HK-2 cells subjected to H/R, the expression of TRAF6 was assessed using Western blot. (D) The cell viability of the H/R + miR-590 inhibitor group was significantly lower than of the H/R + NC inhibitor group by the CCK-8 assay. (E) Western blot analysis of autophagy-related proteins (LC3, p62, Beclin-1) revealed that H/R-induced autophagy was inhibited in the H/R + miR-590 inhibitor group. The data are expressed as the mean ± standard deviation; N = 4–5 for Western blot. ∗P < 0.05 vs. the control group; †P < 0.05 vs. the NC inhibitor group. CCK-8: Cell Counting Kit-8; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; H/R: Hypoxia/reoxygenation; LC3: Microtubule associated protein 1 light chain 3; NC: Negative control; SD: Standard deviation; TRAF6: Tumor necrosis factor receptor-associated factor 6.