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. 2010 May 17;107(24):10783–10790. doi: 10.1073/pnas.0914507107

Fig. 3.

Fig. 3.

Differences in PRC1 binding between TS/XEN cells and ES cells. (A–D) The ability of H3K27me3 to recruit downstream mediators in TS and XEN cells was investigated using ChIP-qPCR on a panel of candidate gene promoters. We included H3K27me3-modified and unmodified promoters that were identified by our genomewide analysis. The panel also contains bivalent promoters classified in ES cells as PRC1-positive (Irx1, Dlx3, Tcfap2a, Lhx2, Npas2, Gbx1, Msx1, Tbx2, Gata6, and Sox17) and PRC1-negative (Pik3r3, Prtg, and Nostrin) (34). Positive (Hoxa9 and Pou5f1) and negative (Gapdh and Kcnq1ot1) control promoters for H3K27me3 were included (61). Ezh2 and Eed binding was detected above background levels at H3K27me3-modified promoters in TS (A4) and XEN (F4) cells. However, binding of the downstream mediator Rnf2 was detected only at low or negligible levels, irrespective of H3K27me3, Ezh2, or Eed status. As we could not identify a promoter that was bound by Rnf2 in TS and XEN cells, we used the Cdx2 promoter in ES (R1) cells as a positive control (indicated by the black bar) (34). Data represent mean plus SD from three biological replicates. Dashed lines indicate 2-fold of mean background levels, as determined using a nonspecific control antibody (background data shown in Fig. S8A).