Figure 1. Dual luciferase reporter gene assays for SMAD4, FRAT1 and BCL2. (A, B, C) Schematic representation of luciferase reporter gene constructs with wild-type miRNA binding sites (3’UTR) and mutated variants (3’UTR mut) for SMAD4, FRAT1 and BCL2. (D, E, F) Dual luciferase reporter gene assays 48 h after co-transfection of HEK293T cells with reporter gene constructs (pMIR-RNL-TK) with the respective wild-type or mutated 3‘UTR for each target and control (pSG5) or miRNA-expression construct (pSG5-miR-34a) in the indicated combinations. Relative luciferase units (RLU) are means ± SEM of three independent experiments performed in duplicates (*, P < 0.05; **, P < 0.01; ***, P < 0.001).