Fig 2: (A) The del4 plasmids, del4_Pu27 (lanes 1-2), del4_Pu27(1(G/A)) (lanes 3-4) and del4_Pu27(5(G/A)) (lanes 5 and 6) were incubated with or without 6xHis-tagged BG4 and pulled down with Co2+ functionalized magnetic beads as indicated in top of the gel-picture. The gel picture shows the result of 20 cycles PCR amplification of pulled down plasmids analyzed in a 1% agarose gel. (B) Linearized del4 plasmids, del4_Pu27 (lanes 1 and 2), del4_Pu27(1(G/A)) (lanes 3 and 4) and del4_Pu27(5(G/A)) (lanes 5 and 6) were incubated with or without TOP1 as indicated in top of the gel picture and subsequently immunoprecipitated with BG4 as described under A. The gel picture shows the result of PCR amplification of pulled down plasmid DNA as described under A. The right panel is a graphical depiction of quantifying the PCR products obtained in five independent experiments. Plotted data represent average ± standard error of the mean, n = 5. * indicates significant difference, * P< 0.05; ** P< 0.005; ns, non-significant difference. Unpaired t-test with Welch's correlation. Band intensities were quantified by using ImageJ.

Fig 3: Quantitative depiction of the Pu27 promoter activity measured in terms of luciferase expression after treatment with equal concentrations of scrambled siRNA (grey bars) or TOP1 targeting siRNA (blue bars). Plotted data represent average +/− standard error of the mean, n = 9. *** indicates significant difference, P< 0.001; ns, non-significant difference. Unpaired t-test with Welch's correlation.