Figure 6.

Modified SOX decoys show enhanced stability and repress reporter gene transactivation. (A) Decoys were incubated in mouse serum and their integrity was assessed by urea-PAGE and SYBR gold nucleic acid staining as a function of incubation time. The ‘0’ lane marks a sample taken immediately after mixing with serum and numbers denote time in hours. UD: undigested decoys. SO: serum only. (B) Decoys were incubated with DNase I and analysed as in (A). (C) The melting of decoys was monitored by heating 1 μM DNA to 90°C at 1°C/min and recording the absorbance at 260 nm. (D) SOX18 exogenously expressed in COS-7 cells activates the expression of a luciferase reporter under control of a regulatory region derived from the VCAM-1 gene (38) in a dose-dependent manner. The inlet illustrates the assay set-up. (E) The effect of decoys at 1000 nM on the VCAM-1 reporter activity was compared 48 h post-transfection using 50 ng of SOX18 plasmid. (F) Effects of the PSCIRC on luciferase expression at different concentrations. (G) Expression of selected genes detected by RT-qPCR in the absence or presence of the PSCIRC. The mean and standard deviation from three experiments each carried out in triplicates is shown. The asterisks (**) denote P < 0.001 (t-test).