Figure 3

DKK3 is a direct target of miR-92b. (A) The putative miR-92b-targeted sequence in the DKK3 gene. TargetScan predicts the binding site to be in the 3′-UTR of DKK3. (B) The DKK3 protein level was assessed 48 h after transfection of U251 and U87 cells with either the miR-92b mimics (100 nM), the control oligonucleotide (100 nM) or the miR-92b inhibitor (100 nM). DKK3 protein levels were detected by western blot assays. β-actin protein was assayed as a control. Scanning densitometry of the blots was used to quantify the Western blotting data. (n=3; means ± SEM **p<0.01, ***p<0.001 t-test). All the data were presented as a representative average of three independent experiments. (C) The U87 cells transfected with the vector containing the DKK3 3'-UTR fragment with the miR-92b binding sequence had inhibited luciferase activity after miR-92b transfection. No luciferase activity change was observed when the cells were transfected with the plasmid containing a DKK3 3′-UTR fragment without the miR-92b binding sites. Each bar represents the mean of three independent experiments. ***p<0.001.