Fig 2: The role of MAX/MGA in recruiting PCGF6-PRC1 to its target genes.(A) Considerable overlap of genes bound by PCGF6 and MAX. Venn diagram depicts the overlap of PCGF6, MAX and MYC target genes in ESCs. Published ChIP-seq data for MAX and MYC were obtained from NCBI GEO (accession number GSM1171650 and GSM1171648, respectively). Odds ratio (OR) and p-values for the overlap between respective groups are indicated. Numbers represent the number of genes bound by each protein or included in each fraction seen in the Venn diagram. (B) Dot plot representation for gene expression changes in Max-KD (left) and Mga-KD (right) ESCs. Expression levels of respective genes (FPKM: determined by RNA-seq) in each group (classified by ChIP-seq) in control (Ctrl) and respective knockdowns (KD) are shown by grey and yellow dots, respectively. The same codes as described in Figure 2A are used. (C) Changes in binding of FLAG-tagged PCGF6, MAX and MGA at the selected targets induced by knockdown of Max or Mga. Local levels of FLAG-tagged PCGF6, MAX and MGA at the respective promoter regions in untreated ESCs or ESCs treated with either control siRNA, Max siRNA or Mga siRNA were determined by ChIP-qPCR. The relative amount of ChIPed DNA is depicted as a percentage of input DNA. Error bars represent standard deviation determined from at least three independent experiments. (D) Changes in local deposition of RING1B, H2AK119ub1, H3K27me3, RYBP and L3MBTL2 at the indicated targets in Max-KO ESCs. Underlined genes are canonical PRC1 targets. Their deposition in Max conditional KO ESCs before (WT) or after doxycycline treatment (Max-KO) was determined by ChIP-qPCR. The relative amount of ChIPed DNA is depicted as a percentage of input DNA. Error bars represent standard deviation determined from at least three independent experiments.DOI: http://dx.doi.org/10.7554/eLife.21064.009