Pho-FLAG and Sce-FLAG binding peaks at PRE2.

<p>(A) A map of the <i>en</i> gene showing the location of the PREs and the probes used for the qPCR (#1–8). (B,C) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048765#s2" target="_blank">Results</a> of X-ChIP experiment with Sce-FLAG (B) and Pho-FLAG (C) driven by <i>en</i>-Gal4 (open bars) or <i>ci</i>-Gal4 (closed bars). Pho binding was also done on all chromatin preparations. The results of a representative experiment are shown. These experiments were done with a different batch of FLAG antibody and different ChIP reagents than those done in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048765#pone-0048765-g005" target="_blank">Fig. 5</a>. Further, 20 larvae were used for each sample instead of 10. Under these conditions, we did not see a difference in the level of binding to the PREs between the “ON” and the “OFF” states; however, the qualitative result, PcG proteins binding to PRE2 in both the “ON” and “OFF” states was the same in these experiments and those in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048765#pone-0048765-g005" target="_blank">Fig. 5</a>.</p>