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. 2019 Dec 12;122(4):578–589. doi: 10.1038/s41416-019-0679-z

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — a The GEPIA database showed that a significant positive correlation between ZEB1 and β-catenin could be observed in CRC tissues. The data from this study also revealed a significant positive correlation between ZEB1 and β-catenin. b The WB analysis showed that the depletion of β-catenin neutralised the promotion of ZEB1 caused by enhanced PLAGL2 expression. (c) Depletion of β-catenin neutralised the promotion of ZEB1 caused by enhanced PLAGL2 expression, which was confirmed by immunofluorescence assays. Scale bars, 100 μm. d The validity of the small-molecule inhibitor PNU-74654 was verified by the WB analysis. e The WB analysis revealed that the PNU-74654, which blocked the interaction between β-catenin and TCF4, thereby neutralising the promotion of ZEB1 caused by enhanced PLAGL2 expression. f, g The ChIP assays were performed to verify the binding between β-catenin/TCF4 complexes and ZEB1 promoter in SW480 and LOVO cells. The bound DNA fragments were amplified by qRT-PCR. Then the products of qRT-PCR were examined by gel electrophoresis on 2% agarose gels. PNU-74654 blocked β-catenin/TCF4 complexes from directly binding to the ZEB1 promoter and neutralised the promotion of ZEB1 caused by enhanced PLAGL2 expression. The data are presented as the mean ± SD from three independent experiments. n.s: no significance. *P < 0.05, **P < 0.01, ***P < 0.001, based on Student’s t-test.