Extended Data Fig. 9: Mutant clones secrete functional BMP ligands.

a–d, Representative in situ hybridization images and quantification of Axin2 (a, b) and Id1 (c, d) on sections of small intestine from Villin-CreERT2;R26R-Confetti or Red2Onco mice 2 w after tamoxifen administration. Arrowheads, fixed mutant crypts. Crypts are marked with dashed outlines. e, f, qPCR analysis of Axin2 (e), and Id1 (f). g, Experimental set-up for h–j. h, Bright-field images of intestinal organoids after 2 days of treatment. The number and size of crypt-like budding structures are reduced in treated organoids. i, qPCR analysis of lineage markers. j, Representative images of LGR5–EGFP organoids show that the number of LGR5⁺ cells decreases following treatment. k, Experimental set-up for l, m. l, m, Bright-field images (l) and quantification (m) of intestinal organoids after 6 days of culture in WENR medium. n, qPCR analysis of BMP ligands (Bmp2 and Bmp7). o, Experimental set-up for p. p, qPCR analysis of Id1 using WT organoids after the CM treatment. q, Bright-field images of intestinal organoids from Villin-CreERT2;R26R-Confetti or Red2Onco mice 1 month after tamoxifen administration. Insets, RFP expression in the mutant organoids. r, qPCR analysis of WT and mutant organoids cultured in ENR medium. In e, f, n, sorted RFP⁺ or YFP⁺ cells from Villin-CreERT2;R26R-Confetti or Red2Onco mice 2 w after tamoxifen administration (4 mg per 20 g body weight, mosaic dosage) were analysed. *P < 0.05, **P < 0.01, ***P < 0.0001; one-way ANOVA with Games-Howell’s multiple comparisons test (b, d) and unpaired two-tailed t-test (e, f, i, m, n, p, r). Quantification graphs show data from three independent experiments (i, m, p, r). Data are mean ± s.d. (b, d, i, m, p) or mean ± s.e.m. (e, f, n, r). For exact P values, see Source Data. Scale bars: 50 μm (a, c, h), 100 μm (j, q) and 500 μm (l).

Source data