Figure 6
DKK2 is a target of miR-27a.
(A) Binding sites for miR-27a in the DKK2 3′UTR were predicted using TargetScan and RNAhybrid software. Red font indicates sequences that were mutated to abolish the interaction between miR-27a and the DKK2 3′UTR. (B) The miR-27a binding site sequences in the DKK2 3′UTR in different species. The miRNA binding sites are shown in grey. Mmu, Mus musculus; Ptr, Pan troglodytes; Mml, Macaca mulatta; Rno, Rattus norvegicus; Cfa, Canis familiaris; Bta, Bos taurus; mdo, Monodelphis domestica; and Oan, Ornithorhynchus anatinus. (C) pmirGLO-DKK2 was co-transfected into CHO cells with miR-27a mimics or NC. Whole cellular lysates were obtained 24 h after transfection and relative luciferase activity was then measured. (D) pmirGLO-DKK2-mut was transfected into CHO cells with miR-27a mimics or NC. (E) Endogenous DKK2 mRNA levels were detected in CHO cells 24 h after transfection with miR-27a mimics or NC mimics and inhibitor or inhibitor NCs. (F) DKK2 protein levels were also monitored using Western blot analysis for 48 h after transfection with miR-27a mimics or mimic NC and inhibitor or inhibitor NCs. **P < 0.01.
