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. 2012 Sep 11;7(9):e44361. doi: 10.1371/journal.pone.0044361

Figure 3. Circular permutation analysis of MA-MADS5 protein using the N10 and c-fos SRE sites.

Figure 3

a Diagrammatic representation of N10 and SRE sites generated by restriction digestion of pAS152 and pAS76 respectively. The location of N10 site and c-fos SRE were indicated by filled boxes. Probes were generated by digestion with MluI, BglII, XhoI, EcoRV, SmaI, StuI, RsaI and BamHI, respectively. b and d Gel mobility shift assay of MA-MADS5 protein bound to each of the circularly permuted probes containing either the N10 or SRE sites (lanes 1–8). DNA-protein complexes were analyzed on a 6% non-denaturing polyacrylamide gel. c and e The data from each circular permutation analysis was shown graphically beneath each set of primary data. The relative mobilities of DNA-protein complexes were normalized for differences in probe mobility and plotted as a function of the position of the center of the N10 or SRE site from the 5′ end of the probe respectively. The points were connected by a curve of the best fit of cosine function. Error bars indicated standard deviations calculated from at least three independent experiments. Representative images from at least three independent experiments have been shown for Figure b and d.