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. 2017 Oct 13;49(10):e385. doi: 10.1038/emm.2017.147

Figure 3.

Figure 3

Upregulated β-catenin in Ctbp2-knockdown ESCs directly targets pluripotency genes. (a) Comparison of co-occupancy regions for Tcf3, stabilized β-catenin and Ctbp2 using published ChIP-seq data.35, 42, 43 In total, 824 regions were co-occupied by Tcf3, stabilized β-catenin and Ctbp2 (A regions), and 726 regions were co-occupied by stabilized β-catenin and Ctbp2 but not Tcf3 (B regions). (b) The mean enrichment levels of β-catenin and Ctbp2 in A regions versus B regions. The mean enrichment levels of β-catenin were higher in A regions than B regions, but the levels of Ctbp2 were similar in both types of regions. (c) Using motif discovery algorithms, Oct4, Sox3, Klf1 and Nanog motifs were predicted with high scores in A regions. (d) The functional analysis of A regions using GREAT showed that enrichment regions were related to epidermis development and stem cell development. (e) Integrated genomics viewer (IGV) represents Ctbp2, stabilized β-catenin, Tcf3, Oct4 and Esrrb in Oct4 and Nanog loci in ESCs. (f) ChIP-qPCR analysis of β-catenin in Ctbp2-occupied Oct4, Nanog, Esrrb, Rex1, Klf2 and NrOb1 loci regions between wild-type and Ctbp2-knockdown ESCs. Values represent the mean±standard deviation (n=3). (g) The TOP-FLASH and FOP-FLASH reporter activity were measured in wild-type, Ctbp2-knockdown and rescued ESCs. Increased β-catenin activity in Ctbp2-knockdown ESCs compared to wild-type ESCs was decreased in rescued ESCs. Value represent mean±standard deviation (n=3). *P⩽0.05, **P⩽0.01.