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.