Extended Data Fig. 6: Mutant p53 promotes balanced growth and differentiation of intestinal organoids.
From: The gut microbiome switches mutant p53 from tumour-suppressive to oncogenic
a, RT–qPCR of Csnk1a1 and Trp53 in jejunal organoids. Mean ± s.e.m. (n, number of mice that were used as a source for organoid cultures) relative to CKIafl/fl (normalized to 1), one-way ANOVA with Tukey’s test. b, e, Immunofluorescent staining of p53 (b) and H&E (e) of jejunal organoids; different levels of dysplasia are evident in e. Nuclear counterstain (immunofluorescence), Hoechst (blue). Scale bars, 100 μm (b) and 50 μm (e). c, d, Bright-field imaging of jejunal (c) and ileal (d) organoids that express p53WT or p53R172H, with or without CKIa knockout induction, and of CKIa/p53 DKO organoids. Inserts show a high-magnification image of a representative organoid. Scale bars, 500 μm. f, RT–qPCR of p53 targets in jejunal organoids. Mean ± s.e.m. (n, number of independent experiments performed with organoid cultures of two different mice), relative to CKIafl/fl (normalized to 1), one-way ANOVA with Tukey’s test. g, Immunofluorescent staining of Ki67 and WNT targets in jejunal organoids. Nuclear counterstain, Hoechst (blue). Scale bars, 100 μm. h, Bright-field imaging and merged immunofluorescence of GFP with p53 or Ki67 in jejunal CKIa/p53 DKO organoids transduced with the indicated lentiviruses. Inserts (bright-field) show a GFP and bright-field merged image of a representative organoid. Nuclear counterstain (immunofluorescence), Hoechst (blue). Scale bars, 500 μm (top); 100 μm (bottom). i, Immunoblot of wild-type and ApcMin/Min organoids. Tubulin, loading control. For gel source data, see Supplementary Fig. 1. Representative data from two or three (b, g–i) or five (c–e) independent experiments.
