Validation of miR-485-5p-YAP1 mRNA Interaction

The interaction between miR-485-5p and YAP1 mRNA was tested by dual luciferase reporter experiment. The YAP1 mRNA 3^′UTR regions containing wide-type (WT) or mutant (MUT) binding site of miR-485-5p was cloned into pmirGLO luciferase reporter (Promega, Madison, WI, USA) to construct YAP1-WT and YAP1-MUT reporter plasmids. The cells were co-transfected with YAP1-WT or YAP1-MUT and miR-485-5p mimic or miR-NC as well as Renilla luciferase (Rluc) control plasmids. After 48 h, the Dual-Luciferase Reporter Assay Kit (Promega, E1910) was used to detect luciferase activity according to the manufacturer’s instructions. The relative luciferase activity was normalized to that of Renilla luciferase.^{45 (link)}