Research Paper Volume 13, Issue 10 pp 14185—14197

Circular RNA ITCH promotes extracellular matrix degradation via activating Wnt/β-catenin signaling in intervertebral disc degeneration

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Figure 5. MiR-17-5p targets SOX4 in NP cells. (A) The interaction of miR-17-5p and SOX4 3’ UTR was identified by bioinformatic analysis using Targetscan (http://www.targetscan.org/vert_72/). (BE) The NP cells were treated with the miR-17-5p mimic or control mimic. (B) The expression levels of miR-17-5p were measured by qPCR in the cells. (C) The luciferase activities of wild type SOX4 (SOX4 WT) and SOX4 with the miR-17-5p-binding site mutant (SOX4 MUT) were determined by luciferase reporter gene assays in the cell. (D) The mRNA expression of SOX4 was analyzed by qPCR in the cells. (E) The protein expression of SOX4 and β-actin was tested by Western blot analysis in the cells. (F) The protein expression of collagen II, aggrecan, MMP13, ADAMTS4, and β-actin was analyzed by Western blot analysis in the NP cells treated with control mimic, miR-17-5p mimic, or co-treated with miR-17-5p mimic and pcDNA3.1-SOX4 overexpression vector. Data are presented as mean ± SD. Statistic significant differences were indicated: * P < 0.05, ** P < 0.01.