Research Paper Volume 11, Issue 15 pp 5705—5725

Figure 6. MiR-27a-3p targets FOXO1 and inhibits its expression. (A) Predicted consequential pairing of the target region (FOXO1 3′UTR) and miR-27a-3p from TargetScan (http://www.targetscan.org/vert_72/). (B) Schematic illustration of the predicted miR-27a-3p binding sites with the 3′UTR of FOXO1. (C) A luciferase activity assay was performed in caki-1 cells co-transfected with miR-27a-3p and the luciferase reporter plasmids driven by either WT or MUT 3′UTR of FOXO1. (D) The expression of FOXO1 at the mRNA level was determined by qRT-PCR analysis in 786-O and caki-1 cells transfected with NC, miR-27a-3p, or miR-27a-3p inhibitor. (E) The expression of FOXO1 at the protein level was determined by western blot analysis in 786-O and caki-1 cells transfected with NC, miR-27a-3p, or miR-27a-3p inhibitor. Three independent experiments were performed and data shown are mean ± SD. Statistically significant differences are indicated as *, P<0.05, **, P<0.01; ns, no significance; Student’s t-test among two groups; ANOVA among multiple groups. miR-27a-3p, microRNA-27a-3p; FOXO1, Forkhead Box Protein O1; UTR, untranslated region; WT, wild type; MUT, mutant type; qRT-PCR, quantitative real-time polymerase chain reaction; NC, negative control; SD, standard deviation.