Fig. 5

Results of prediction and validation of the target relationships between miRNAs and the 3’UTR of the AANAT gene. The target sites of miR-89 (a) and miR-201 (b) in the 3’UTR of the AANAT gene were predicted by RNAhybrid software. The results of validation of the target relationships of miR-89 and miR-201 through the dual luciferase reporter system are shown in (c) and (D). WT + miR-89: cells cotransfected with pmirGLO-AANAT wild-type 3’UTR (0.025 μg) and pcDNA6.2-GW/miRNA-89 (0.075 μg); WT + NE: cells cotransfected with pmirGLO-AANAT wild-type 3’UTR (0.025 μg) and pcDNA6.2-GW empty vector (0.075 μg); MUT + miR-89: cells cotransfected with pmirGLO-AANAT mutant-type 3’UTR (0.025 μg) and pcDNA6.2-GW/miRNA-89 (0.075 μg); MUT + NE: cells cotransfected with pmirGLO-AANAT mutant-type 3’UTR (0.025 μg) and pcDNA6.2-GW empty vector (0.075 μg); blank: cells without transfected vector. The test groups for miR-201 were similar to those for miR-89. **: P < 0.01