Fig. 3: Corticosterone reproduces the effect of stress on ISCs.

a Schematic representation of CORT administration. C57BL/6 mice were injected intraperitoneally with CORT (5 mg/kg) or vehicle once daily for 3 weeks at varying times of the day. CORT corticosterone. b Plasma CORT levels in mice measured 3 h after a single vehicle or CORT injection (n = 4). c, d H&E staining (c) and quantification of crypt depth (d) in the jejuna of the vehicle (n = 4) and CORT-treated (n = 5) mice. Scale bar: 50 µm. e, f Ki-67 staining (e) and quantification of Ki-67⁺ cells per crypt (f) in the jejuna of the vehicle (n = 4)- and CORT-treated (n = 5) mice. Scale bar: 20 µm. g, h Representative flow cytometric plots showing EGFP⁺ cells (g) and quantification of the percentage of ISCs (Lgr5-EGFP^High) and progenitor cells (Lgr5-EGFP^Low) (h) in the small intestine of the vehicle- and CORT-treated Lgr5-EGFP-IRES-creERT2 mice (n = 6). i, j Olfm4 staining (i) and quantification of Olfm4⁺ cells per crypt (j) in the jejuna of the vehicle (n = 4)- and CORT (n = 5)-treated mice. Scale bar: 20 µm. k‒m EdU and Olfm4 co-staining (k), quantification of EdU⁺ cells per crypt (l) and the percentage of EdU⁺ Olfm4⁺ cells (m) in the jejuna of the vehicle- (n = 4) and CORT (n = 5)-treated mice. EdU (0.2 mg/25 g body weight) was administered to mice 1.5 h prior to sacrifice. Scale bar: 20 µm. n‒q Representative images of day 4 organoids (n) and quantification of organoid formation (o), bud number (p) and surface area (q) from the vehicle- and CORT-treated mice (n = 3). Scale bar: 100 µm. The data are presented as the mean ± SD. The statistical analysis was performed by an unpaired two-tailed Student’s t-test for normally distributed data or the Mann‒Whitney test for non-normally distributed data. ns, p ≥ 0.05, *p < 0.05, **p < 0.01, ***p < 0.001.