Fig. 5.

Inhibitory effect of the aptamer to ER-mediated transcriptional activation in MCF7 cells. (A) Accumulation level in MCF7 of AptER-1×2 expressed from the pSUPER vector. Cells are harvested at several time points within 96 hours post-transfection of the pSUPER/AptER-1×2 plasmid. The level of AptER-1×2 is quantified using real-time RT-PCR and expressed as relative expression level (fold) of β-actin. (B) Dual-luciferase assays for aptamer expressed using two different vectors. MCF7 cells are co-transfected with the firefly luciferase vector (3×ERE TATA luc), Renilla luciferase vector and an aptamer expression vector of interest. “Ctrl” is a scrambled RNA expression vector used in place of the aptamer expression vectors. All experiments are carried out in triplicate and the average is used to make the plot. (C) Specificity of the inhibitory effect. 3×ERE TATA luc is replaced with firefly luciferase reporter gene 2×PRE TK luc or pGL3 luc. The pSUPER vectors are used to express aptamers or the control RNA. Each experiment was repeated at least three times and the standard deviation was shown as error bars.