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. 2019 Dec 5;9:18440. doi: 10.1038/s41598-019-54890-9

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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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β-catenin mutation affects the localization and activation of β-catenin, as well as morphology of HCT116 cells. (a) Morphologies of HCT116 cells containing a wild-type (WT) β-catenin allele (HCT116-WT, WT), a mutant β-catenin allele (HCT116-MT, MT), or both wild-type and mutant alleles (HCT116-P, Parent). Red and blue arrows indicate mesenchymal-like and epithelial-like morphologies, respectively. (b) Relative β-catenin mRNA expression levels in HCT116 cell lines determined by qRT-PCR. (c) Western blot analysis of β-catenin protein levels in HCT116 cell lines. (d) Quantification of band intensities for the western blot shown in (c). (e) Top panel: western blot analysis of β-catenin levels in HCT116 cell lines subjected to cell fractionation. Bottom panel: quantification of band intensities for the western blot shown at top. hnRNPC and α-tubulin were used as nuclear and cytoplasmic markers, respectively. (f) Immunofluorescence microscopy analysis of β-catenin (stained in green) in HCT116 cells; DAPI staining is shown in blue. (g) Activation of WNT/β-catenin pathway in HCT116 cells, measured by TOP/FOP luciferase reporter assay. Error bars represent the standard deviation (SD) of the mean of two independent experiments. All assays shown in (c–f) were carried out in triplicate.