Extended Data Fig. 1: Intestinal epithelial cell profiling by flow cytometry, Irf2 expression in steady state WT mice, and apoptosis and Wnt/Notch signalling in mucositis-induced Irf2−/− mice.
a, Flow cytometry gating strategy for ISCs and Paneth cells in Lgr5-EGFP-Ires-CreERT2 mice. ISCs and Paneth cells were isolated as Lgr5hi cells and CD24hiUEA1hi cells within EpCAMhi crypt ECs, respectively. b, Representative images of Irf2 mRNA detection in the jejunum epithelium of WT mice using RNAscope in situ hybridization assays. 3 mice. Irf2 mRNA is depicted as brown dots. Most epithelial cells, including ISCs (red arrowheads), transit amplifying cells (red dashed lines) and goblet cells (Gob, black arrowheads), expressed Irf2 mRNA. Orange arrows indicate Irf2 mRNA dots in ISCs. c-e, Gene expression of Olfm4 (c), Lyz1 (d) and Irf2 (e) in FCM-sorted ISCs and in Paneth cells relative to total crypt ECs. Data represent means ± s.d. of n=3 mice. Statistics were determined by one-way ANOVA followed by Tukey’s multiple comparisons test. f, Representative images of TUNEL staining in the jejunum of Irf2+/− and of Irf2−/− mice injected with 5-FU as in Fig. 1. Intestines were harvested on d 2 and 6 (Irf2+/−, naïve, 3 mice; Irf2+/−, day 2, 3 mice; Irf2+/−, day 6, 4 mice; Irf2–/−, naïve, 3 mice; Irf2–/−, day 2, 3 mice; Irf2–/−, day 6, 4 mice). g, Gene expression of Olfm4, Ascl2 and Hes1 in the jejunum of Irf2−/− mice compared with Irf2+/− mice on day 6 after 5-FU injection. Data represent means ± s.d. of n=4 mice. Statistics were determined by two-tailed Mann-Whitney test. Scale bars: 20 μm (b) and 100 μm (f).
