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. 2018 Mar 19;9:1132. doi: 10.1038/s41467-018-03426-2

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© The Author(s) 2018

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. 4 — Reduced Wnt ligand secretion does not affect Apc-deficient cell growth but accelerates tumorigenesis. aVilCre^ERApc^fl/fl mice were induced and treated with LGK974 starting the following day. Mice were sampled at day 4 post induction, and proliferation assessed by BrdU staining. Scale bar = 50 µm. bLgr5Cre^ERApc^fl/fl mice induced with 0.15 mg tamoxifen and treated with LGK974/vehicle 1 day p.i. The number of macroscopic adenomas was scored when mice showed signs of intestinal adenomas or at 100/130 day timepoint. Each dot represents one mouse, untreated N = 5, vehicle N = 7 and LGK974 N = 9, black dot = mean, error bars = s.e.m., Mann–Whitney U test: untreated vs. LGK974 p = 0.002639, vehicle vs. LGK974 p = 0.0006014. c Immunohistochemistry for β-catenin showed only small microadenomas in the vehicle group, in contrast to adenomas found after LGK974 treatment. Scale bar = 100 µm. d Example image of Lgr5Cre^ERApc^fl/fltdTom^fl/+ mouse 10 days post induction (3 mg, LGK974 treatment). Immunohistochemistry for tdTomato (RFP) shows fully labelled crypt. Arrow marks unlabelled cell (probably Paneth cell), suggesting a newly labelled crypt. RNA in situ for Apc exon 14 shows that only half of the crypt has recombined (dashed area). Scale bar = 50 µm. eLgr5CreER Apc^fl/fl mice were induced with 3 mg tamoxifen and treated with LGK974/vehicle starting at day 1 p.i. Crypts were scored based on immunohistochemistry for β-catenin and categorised into partial and full crypts. The ratio of full crypts is in relation to the sum of full and partial crypts. f Scoring of partial and full crypts at different timpoints in absolute numbers. N = 3 mice for each timepoint and each group, error bars = s.e.m. gLgr5CreER Apc^fl/fl mice induced with 3 mg tamoxifen were treated with LGK974 or vehicle starting at day 1 p.i. Mice were sampled when signs of intestinal tumour burden were apparent. Untreated N = 5, vehicle (VEH) N = 7, LGK974 N = 9 mice, log-rank test: vehicle vs. LGK974 p = 0.00019, untreated vs. LGK974 p = 0.0222. h Number of adenomas scored on histological sections. Untreated N = 5, vehicle (VEH) N = 6, LGK974 N = 9 mice. Each dot represents number of lesions per mouse; box indicates mean ± standard deviation. Mann–Whitney U test for LGK974 vs. control mice (untreated and vehicle) p = 0.02496