FIGURE 2. miRNAs directly target sequences in the IFNβ 3′UTR.
miRNA mimics reduce luciferase expression from an IFNβ 3′UTR-containing dual firefly/Renilla luciferase reporter vector (A). Normalized luciferase expression is reduced in HEK-293T cells transfected with specific miRNA mimics and equimolar mixtures of the four indicated miRNAs, but not in cells transfected with a control miRNA (CmiR). Each condition is presented relative to a no-miRNA control, set equal to one. Error bars are SEM from three independent experiments. Individual predicted IFNβ miRNA recognition elements cloned downstream of GFP suppressed GFP expression in HEK-293T cells, but three of four mutated MREs relieved this suppression (B). Fluorescence intensity is displayed as the ratio of mutant to wild-type MRE, normalized to red fluorescence (from pdsRed transfection control). Error bars represent SEM.
