Figure 6 of Silva, Mol Vis 2011; 17:2228-2240.


Figure 6. Luciferase reporter assay. A: The firefly luciferase reporter construct (pISORAX plasmid) contains a partial-length 3′-UTR of RAX mRNA in the 5′ to 3′ orientation with respect to the firefly-luc open reading frame with the firefly luciferase start and stop codons and the potential miR-29 target site. The pISORAX plasmid contains the partial-length 3′-UTR of RAX mRNA in the 5′ to 3′ orientation with respect to the firefly-luc open reading frame. The firefly luciferase start and stop codons and the potential miR-29 target site are also shown. HEK293 cells were co-transfected with pISO (B) or pISORAX (C) and pRL-TK together with a synthetic miR-29b mimic or dsRNA as a negative control. The indicated concentrations refer to the quantity of miR-29b mimic or dsRNA used for the transfection. The luciferase activity of firefly-luc and Renilla-luc were measured at 24 h and 48 h after transfection. The firefly-luc activity was normalized to Renilla-luc expression. Data are means±standard error of the mean from three independent experiments, each containing three replicates.