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. 2016 Oct 15;7(48):79372–79387. doi: 10.18632/oncotarget.12676

Figure 2. miRNAs regulate MYCN expression through direct and indirect pathways.

Figure 2

A. IPA analysis identifies the potential mechanisms by which the differentiation-inducing miRNAs regulate MYCN expression. B. The predicted interactions of miRNAs with the targets sites in the 3′UTR of MYCN mRNA. The seed sequences are underlined. C-E. Validation of the target sites of the miRNAs in the 3′UTRs of MYCN mRNA by luciferase reporter assay in BE(2)-C, KELLY and SKNSH cells. Cells were co-transfected with the luciferase reporter vector expressing either the wildtype 3′UTR of MYCN (WT 3′UTR) or mutated MYCN 3′UTR (mutant 3′UTR) and miRNA mimics. After 72 h of transfection, cells were lysed and luciferase activity was measured. Shown are normalized luciferase activities with the luciferase activity associated with the WT 3′UTR normalized to those associated with the corresponding mutant 3′UTRs. *, p<0.05 compared to the mutant 3′UTR.