Skip to main content
. 2017 Dec 9;8(2):491–503. doi: 10.1534/g3.117.300311

Figure 1.

Figure 1

β-catenin regulates known PRC2 targets and can physically interact with PRC2. (A) Schematic demonstrating the isolation of CM+ectoderm by manual dissection. The isolated tissue is comprised of the supraorbital CM and overlying ectoderm. (B) Gene ontology analysis of differentially expressed genes in E13.5 En1Cre/+;R26R/+;β-cateninfl/+ control (n = 5) and En1Cre/+;R26R/+;β-cateninfl/∆ mutant CM+ectoderm (n = 4). Overlapping gene sets from MsigDB Perturbations ontology ranked by binomial P-value, which accounts for varying sizes of gene regulatory domains, from GREAT. Green bars represent gene sets associated with PRC2. The ranked list is as follows: (1) Genes with high-CpG-density promoters bearing H3K4me2 and H3K27me3 in the brain (MsigDB M1941). (2) Genes with H3K27me3 on their promoters in human embryonic stem cells identified by ChIP on chip (MsigDB M10371). (3) Genes identified as targets of SUZ12 in human embryonic stem cells by ChIP on chip (MsigDB M2291). (4) Genes identified as targets of EED in human embryonic stem cells by ChIP on chip (MsigDB M2736). (5) Genes coordinately upregulated in a compendium of adult tissue stem cells (M1999). (6) Genes upregulated in uterus upon knockout of BMP2 in the uterus (MsigDB M2324). (7) Genes downregulated in mouse embryonic stem cells upon deletion of SUZ12 (MsigDB M2291). (8) Genes upregulated in human lung fibroblasts (IMR90) after knockdown of RB1 by RNAi (MsigDB M2129). (9) Genes downregulated in breast cancer (MsigDB M13072). (10) Genes downregulated in immortalized nontransformed mammary epithelium (HMLE) after knockdown by RNAi or expression of a dominant negative form of CDH1 (MsigDB M11790). (C) Integrated Genome Viewer representation of H3K27me3 ChIP-sequencing on E13.5 En1Cre/+;R26R/+;β-cateninfl/+ control CM+ectoderm of Sox9 (chr11:112,641,524–112,651,071), Col2a1 (chr15:97,804,033–97,837,155), Col9a2 (chr4:120,710,171–120,729,930), and Col11a2 (chr17:34,174,382–34,205,187). (D) RT-qPCR of enriched CM after bead purification with an antibody specific to PDGFRα. A reduction in K14 expression levels demonstrates purification of the mesenchyme. (E) Co-immunoprecipitation of β-catenin with components of PRC2. CM+ectoderm was isolated by manual dissection and the mesenchyme was purified using a PDGFRα antibody. IgG was used as a negative immunoprecipitation control. ChIP, chromatin immunoprecipitation; chr, chromosome; CM, cranial mesenchyme; MsigDB, Molecular Signatures Database; PRC2, Polycomb Repressive Complex 2; RNAi, RNA interference; RT-qPCR, reverse transcriptase-quantitative polymerase chain reaction.