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. 2021 Jan 4;12:56. doi: 10.1038/s41467-020-20255-4

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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. 3 — a H&E, BrdU incorporation and RNAscope for Lgr5 and Olfm4 on Vil-CreER^T2 Apc^fl/fl (APC), Vil-CreER^T2 Apc^fl/fl, Vav2^−/−, Vav3^−/− (APC V2V3) and Vil-CreER^T2 Apc^fl/fl Tiam1^−/− (APC T). Scale bar represents 100 μm for H&E and BrdU and 50 μm for RNAscope images. b Quantification of BrdU positive cells. N = 6, 5 and 6 biologically independent animals for Vil-CreER^T2 Apc^fl/fl (APC), Vil-CreER^T2 Apc^fl/fl, Vav2^−/−, Vav3^−/− (APC V2V3; p = 0. 0303 as determined by a two-tailed Mann–Whitney) and Vil-CreER^T2 Apc^fl/fl Tiam1^−/− (APC T; *p = 0.0260, as determined by a two-tailed Mann–Whitney) respectively. Data are presented as mean values ±SD. c Quantification of clonogenicity assay of intestinal organoids. N = 6, 3 and 3 biologically independent animals for Vil-CreER^T2 Apc^fl/fl (APC), Vil-CreER^T2 Apc^fl/fl, Vav2^−/−, Vav3^−/− (APC V2V3; p = 0.5476, as determined by a two-tailed Mann–Whitney) and Vil-CreER^T2 Apc^fl/fl Tiam1^−/− (APC T) respectively (p = 0.5476, as determined by a two-tailed Mann–Whitney). Data are presented as mean values ±SD. d, e Survival of Lgr5-EGFP-IRES-creER^T2 Apc^fl/fl (Lgr5 APC), Lgr5-EGFP-IRES-creER^T2 Apc^fl/fl Vav2^−/−, Vav3^−/− (Lgr5 APC V2V3) and Lgr5-EGFP-IRES-creER^T2 Apc^fl/fl Tiam1^−/− (Lgr5 APC T). d N = 19 and 18 biologically independent animals for Lgr5 APC and Lgr5 APC V2V3 respectively (p = 0.7142, as determined by Log-rank (Mantel-Cox) test). e N = 15 and 10 biologically independent animals for Lgr5 APC and Lgr5 APC T respectively (under the control of Lgr5-EGFP-IRES-creER^T2). (p = 0.1982, as determined by Log-rank (Two-tailed Mantel-Cox test). The same Lgr5 APC control cohort was used in both d and e as well as in Fig. 4f and Supplementary Fig. 6D. f Quantification of intestinal tumour burden following APC loss in the Lgr5 stem cell compartment. Tumour burden is determined as percentage of intestine which is covered by lesion or adenomas. Lgr5-EGFP-IRES-creER^T2 Apc^fl/fl (APC; n = 9 biologically independent animals) vs Lgr5-EGFP-IRES-creER^T2 Apc^fl/fl Vav2^−/−, Vav3^−/− (APC V2V3; n = 6 biologically independent animals) p = >0.9999 as determined by a two-tailed Kruskal–Wallis with Dunn’s multiple comparisons test. Lgr5-EGFP-IRES-creER^T2 Apc^fl/fl (APC) vs Lgr5-EGFP-IRES-creER^T2 Apc^fl/fl Tiam1^−/− (APC T; n = 7 biologically independent animals), p = 0.7532 as determined by a two-tailed Kruskal–Wallis with Dunn’s multiple comparisons test. Data are presented as mean values ±SD. g Solid adenomas (H&E) were observed in each genotype, indicated by the arrow. Scale bar represents 500 μm.