Extended Data Fig. 7: Genetic ablation of Yap prevents spheroid formation and Sca-1+ stem-cell expansion in fibroblast–crypt organotypic co-cultures.
From: Paracrine orchestration of intestinal tumorigenesis by a mesenchymal niche
a, Crypts isolated from the small intestines of Yap1f/f and Yap1ΔIEC mice were grown into organoids by 3D culture with OGM supplemented with 0.5 mg ml−1 recombinant mouse epiregulin (Ereg) as previously described18, or in a co-culture with wild-type primary mouse intestinal fibroblasts with OGM without Ereg supplementation. Indicative images and quantification of the percentages of crypts, organoids and spheroids grown per 3D structure are shown. n = 2 cultures per condition. Data are representative of two independent experiments. Scale bars, 100 μm. Two-way ANOVA. Data represent mean ± s.e.m. ****P < 0.0001. b, Intestinal crypts isolated from the small intestines of Yap1f/f and Yap1ΔIEC mice were co-cultured with wild-type primary mouse intestinal fibroblasts. On day 4, these co-cultures and control Yap1f/f organoid cultures were processed into single-cell suspensions and analysed by flow cytometry for Sca-1 expression in Cd24+ epithelial cells. n = 2 cultures per condition. Scale bars, 100 μm. FSC, forward scatter. Unpaired two-tailed t-test. Mean ± s.e.m. ***P < 0.001.
