DNM3OS Enhances the Apoptosis and Senescence of Spermatogonia Associated with Nonobstructive Azoospermia by Providing miR-214-5p and Decreasing E2F2 Expression

Figure 4

DNM3OS negatively regulated E2F2 expression by competitively binding to miR-214-5 p. (a) The TargetScan, microT, and miRmap databases were used to create a Venn diagram, and the intersection predicted 11 overlapping target genes of miR-214-5p. (b) Schematic representation of miR-214-5p target sites in the 3^′-UTR of E2F2 mRNA and the 3^′-UTR of the E2F2 mutant mRNA containing altered nucleotides in the putative target site (E2F2-3^′-UTR-MUT). (c) MiR-214-5p mimics or NC mimics were simultaneously transfected into GC-1 spg cells containing the wild-type or mutant E2F2-3^′-UTR, and luciferase reporter gene assays were performed to measure luciferase activity in the cells. represents NC mimics vs. miR-214-5 p mimics; (d) and (e) E2F2 expression at the mRNA and protein levels was determined in GC-1 spg cells that had been transfected with si-DNM3OS, and then incubated for 24 hr with 0.5 μM ADR. represents si-NC vs. si-NC + ADR; ^### represents si-NC + ADR vs. si-DNM3OS + ADR; data represent the mean value ± SD.

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