fig4

Figure 4. Most val-miR218 targets have reduced mRNA and protein expression after val-miR218 overexpression. (A) Val-miR218 target gene expression after val-miR218 overexpression in U2OS cells. The relative mRNA expression of val-miR218 treated cells was compared to those treated with NC miRNA. The relative mRNA expression of NC miRNA treated group was set as 1. n = 3. (B) Representative immunoblots of U2OS cells transfected with val-miR218 or NC miRNA. GAPDH and β-actin are housekeeping genes. The protein of interest was first adjusted to housekeeping proteins. Then, the relative protein expression of val-miR218 treated cells was compared to those treated with NC miRNA. The relative protein expression of NC miRNA treated group was set as 1. n = 4. (C) Dual-luciferase assay of 13 randomly selected miRNA recognition elements (MRE). Normalized to the luciferase activity of cells transfected with the corresponding vector only. n = 3. Compared cells treated with NC miRNA, * P < 0.05.