Extended Data Fig. 11: Functional validation of oncogene-driven niche remodelling.
From: Tracing oncogene-driven remodelling of the intestinal stem cell niche
a, b, qPCR analysis of Id1 (a), Axin2 and Lgr5 (b) after administration of indicated inhibitor. c, Fraction of monoclonal WT (YFP+) crypts remote from (Remote) or proximate to (Prox) mutant crypts in Red2Onco mice. d, Heat maps indicate relative clone fractions of the indicated sizes. Data are mean ± s.e.m. e, Fraction of monoclonal (RFP+) mutant crypts in Red2Onco mice. f, g, Representative confocal images (f) and quantification (g) of EGFP+ (LGR5+) ISCs. Images are representative of tissues quantified in g. Arrows, proximate WT crypts; arrowheads, fixed mutant crypts. h, Representative confocal images of whole-mount small intestine. Images are representative of tissues quantified in i. Arrowheads, fixed mutant crypts. i, Violin plots of proximate WT crypt size. j, k, RNA in situ hybridization (j) and quantification (k) of Bmp2. Arrowheads, fixed mutant crypts. l, m, Representative multiplexed in situ hybridization images (l) and quantification (m) of Rspo3 in Cd81+ cells. Arrow: Cd81-positive STC2 cells; Arrowheads, fixed mutant crypts. Whole mount (f–i) and sections (j–m) of small intestine from Lgr5-EGFP-IRES-CreERT2 control (L5), Lgr5-EGFP-IRES-CreERT2;LSL-KrasG12D (enKR) or PIK3CALat-H1047R (enP3) mice 2 w after tamoxifen administration. In c–e, graphs show data collected 2 w after concomitant administration of indicated drug and tamoxifen. Crypt borders are marked by dashed outlines (f, h, j, l). In f, h, j, l, white (f, h) or red (j, l): immunostaining for mutant KRAS(G12D) in enKR, or p-AKT in enP3. *P < 0.05, **P < 0.01, ***P < 0.0001; one-way ANOVA with Games-Howell’s multiple comparisons test (k, m) and unpaired two-tailed t-test (c, e, g, i). Data are mean ± s.d. (a, b, g, k, m) or mean ± s.e.m. (c, e). For exact P values, see Source Data. Scale bars: 50 μm (f, h, j) and 25 μm (l).
