Fig. 5: miR-802 downregulates Ran expression by directly binding its 3’-UTR.

a Schematic diagram of miR-802 identification. Bioinformatics algorithms were employed to identify potential regulators. We used the TargetScan miRNA database to computationally predict 123 potential miRNAs, 34 of which target the Ran 3’-UTR and are conserved among vertebrates. Among these 34 miRNAs, we focussed on miR-802, miR-40-5p and miR-142-3p because they rank among the top five in both the total context ++ score and aggregate P_CT algorithms and might play a negative role in tumorigenesis according to previous studies. b Western blot analysis of Ran expression in HCT116 cells transfected with miR-802, miR-40-5p or miR-142-3p mimics and their controls. c qRT-PCR analysis of Ran expression (above) and miR-802 expression (below) in a panel of CRC cells. d Ran expression (above) and miR-802 expression (below) in 12 cases of primary CRC tissues and paired normal tissues detected by qRT-PCR. e Western blot and qRT-PCR analysis of Ran expression in HCT116 (left) and DLD-1 (right) cells transfected with miR-802 mimic or negative control and in HCT116 cells transfected with miR-802 mimic or negative control (**P < 0.01). f Left: a schematic representation of the predicted duplex sequences between the Ran 3’-UTR and miR-802. Mutations were designed at the predicted miR-802-binding sites (middle and right). The Ran wild-type and mutant reporter plasmids were co-transfected with a miR-802 mimic (middle), miR-802 inhibitor (right) or the respective controls. Luciferase activity values were measured and analysed (n.s.: not significant; *P < 0.05, **P < 0.01).