Figure 4. A. VEGFA was a direct target of miR-150-5p in CRC. (A) The direct target genes of miR-150-5p were predicted using the PicTarSites, miRandaSites and Tarbase databases. (B) Wild-type and mutant VEGFA-3’UTR sequences were cloned into luciferase reporter. Luciferase activity was determined in HCT116 and 293T cells cotransfected with agomiR-150-5p or agomiR-NC and pmirGLO-VEGFA-3’UTR-WT or pmirGLO-VEGFA-3’UTR-Mut. Luciferase activities were normalized to that of renilla luciferase. C, D. qRT-PCR (C) and western blot (D) analyses showed that both VEGFA mRNA and protein expression levels were dramatically suppressed by agomiR-150-5p in HCT116 and HCT8 cells, GAPDH was used as the internal control. **p<0.01.