Fig. 6. The engagement of PRC1 and PRC2 is gradually decreased upon Gata6-mediated differentiation of ES cells into primitive endoderm lineages. (A) Western blot analysis demonstrating changes in the levels of Oct3/4, Ring1B, Phc1, Ezh2, Eed, Suz12 and H3K27me3 after conditional activation of Gata6 by dexamethasone (Dex) treatment of G6GR ES cells. Lamin B and CBB staining confirmed equal loading. (B) Significant overlap of expression profiles in Ring1A/B-dKO (day 4), Oct3/4-KO (day 1) and Gata6-differentiated (day 2) ES cells. Pearson's correlation of expression changes for probes that belong to specific GO classifications are shown by bars. Functional groupings are indicated above each graph. Error bars represent 95% confidence intervals of correlation coefficients calculated by Z transformation. (C) ChIP analysis showing binding of Ring1B, Phc1 and Eed at the promoter regions of the selected target genes after Dex treatment in G6GR ES cells. The relative amount of immunoprecipitated DNA is depicted as a percentage of input. Error bars represent s.d. determined from at least three independent experiments.