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. 2021 May 5;12(5):449. doi: 10.1038/s41419-021-03733-5

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© The Author(s) 2021

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. 2 — A Overexpression or knockdown of β-catenin was conducted in SW480 and SW620 cells transfected with SOX2 clone or SOX2 shRNA, then immunoblot was performed with indicated antibodies. B–F The ability of chemoresistance (B), proliferation (C), stemness (D), migration (E), and invasion (F) in SW480 cells co-transfected with SOX2 clone and β-catenin siRNA was assessed by drug sensitivity assay, cell viability assay, tumor sphere formation assay, wound healing assay and transwell invasion assay. G β-catenin is a potential binding partner of SOX2 predicted by the STRING database. H Immunofluorescence was performed to analyze the co-localization of SOX2 and β-catenin in SW480 and SW620 cells. I co-IP assay was used to analyze the interaction between SOX2 and β-catenin in both cytoplasm and a nuclear fraction of SW480 and SW620 cells. J Western blot detected the expression of SOX2 and β-catenin in the nucleus and cytoplasm of SOX2-overexpressing SW480 cells and SOX2-silencing SW620 cells. K TOP/FOP Flash reporter assay was used to detected β-catenin transcriptional activity in SW480 and SW620 cells transfected with SOX2 clone or SOX2 shRNA. L, M Total expression of β-catenin and its target genes (c-Myc, cyclin D1, and Axin2) was detected by qRT-PCR (L) and western blot (M) in SOX2-overexpressing SW480 cells and SOX2-silencing SW620 cells. N Effect of β-catenin overexpression or knockdown on SOX2 expression was detected by western blot in SW480 and SW620 cells. O Effect of β-catenin overexpression or knockdown on SOX2-mediated ABCC2 promoter transactivation was assessed in HEK293T cells by luciferase reporter assay. Experiments were conducted in triplicate. Data are shown as mean ± SEM. *P < 0.05, **P < 0.01, and ***P < 0.001.