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. 2020 Jan 23;53(2):e12751. doi: 10.1111/cpr.12751

Figure 3.

Figure 3

CCND1, DNMT3B and NOP14 were identified as direct downstream targets of miR‐502‐5p. A, Bioinformatic prediction analysis. MiRWalk and TargetScan online databases were used to predict potential downstream targets of miR‐502‐5p. B, RT‐qPCR assay. Ten candidate targets were selected, and the mRNA levels of these genes were measured. Only CCND1, DNMT3B and NOP14 were significantly downregulated in both cell lines. C, KEGG pathways analysis. KOBAS tool was used to analyse the pathways involved (representative pathways are presented). D, Dual‐luciferase reporter assay. The luciferase activity was significantly reduced after miR‐502‐5p mimics were transfected into the wild‐type group. However, no marked changes in the luciferase activity were observed in the mutated type group. E, Western blot assay. Protein levels of several candidate targets were measured after transfection with miR‐502‐5p mimics. F, Schematic diagram of the miR‐502‐5p–targeting region of CCND1, DNMT3B and NOP14 with seed matching. Error bars represent the SE obtained from three independent experiments; *P < .05