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. 2023 May 19;14:2861. doi: 10.1038/s41467-023-38581-8

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© The Author(s) 2023

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. 7 — a qPCR analysis of LRP5, LRP6, AXIN2, CCND1, CTGF, CYR61, ALDH1A1 and NANOG in the indicated treatment groups in GX006 parental and GX006 5FU + CDDP resistant organoids. b Representative immunofluorescence images and quantification of β-catenin staining in the indicated treatment groups in GX006 parental and GX006 5FU + CDDP resistant organoids. Scale bar, 50μm (low magnification) and 20μm (high magnification). c Western blot analysis for expression of LRP5, LRP6, β-catenin, AXIN2, cyclin D1 in the indicated treatment groups in GX006 parental and GX006 5FU + CDDP resistant organoids. β-actin is used as the loading control. d Representative immunofluorescence images of GX006 parental and GX006 5FU + CDDP resistant xenografts stained with β-catenin. Scale bar, 20μm (low magnification) and 10μm (high magnification). e Western blot for β-catenin following cycloheximide (CHX) treatment for 0, 2, 4 and 8 hours in GX006 parental, GX006 5FU + CDDP resistant organoids and GX006 5FU + CDDP resistant organoids treated with SSI4. (a, c, e) n = 3 independent experiments; (b) n = 3 independent experiments. (d) n = 15–19 randomly captured field of view. Parental-DMSO and Resistant-COMBO, n = 15 images; Parental-5FU + CDDP and Resistant-DMSO, n = 17 images; Resistant-5FU + CDDP, n = 16 images; Resistant-SSI4 and, n = 19 images. Significance were calculated by (a, b) two-way ANOVA; (d, e) one-way ANOVA. All data were presented as mean ± standard deviation. COMBO for combination, MFI for mean fluorescence intensity. ns for not significant. Source data are provided as a Source Data file.