2.9. Dual Luciferase Reporter Assay

The Keap1 gene dual luciferase reporter was constructed with either wild-type or mutant promoter. After cotransfection of miR-24-3p mimic or miR-24-3p inhibitor with these reporter vectors along with the renin expression plasmid into H9C2 cells, we detected and analyzed the luciferase activity using the luciferase reporter assay kit. To analyze the data, we designated firefly luciferase activity as M1 and Renilla luciferase activity as M2, and we calculated the ratio of M1 to M2. The relative activity of the reporter gene was compared between each group to determine the target of miRNA.