Figure 4

Validation of miR-146a target genes in the Renilla luciferase reporter system. Seven potential miR-146a target genes predicted by the miRanda algorithm were chosen for validation. For each predicted target gene a luciferase reporter vector was constructed in which the predicted miR-146a binding site was cloned into the 3’ UTR of a Renilla luciferase reporter gene. The Renilla luciferase activities were normalized to Firefly luciferase activities (under the control of an independent promoter). The relative expression of each Renilla luciferase target construct was compared between cells expressing miR-146a and those expressing the scrambled control sequence (SC) using a t-test for statistical significance (p < 0.05). Error bars indicate standard deviation. CASP6, a gene containing no binding site for miR-146a but predicted to contain a miR-143 target site was used as negative control.