Figure 1.

Impaired Intestinal Regeneration and Organoid Formation in TX-Treated Villin-CreER^T2 Mice

(A–O) Villin-CreER^T2 mice were treated with TX (100 mg/kg) or vehicle (V; also VEH) daily for 5 days, and 1 day later either (A–O) challenged with 12 Gy γ-irradiation or (P–S) tested for organoid-forming efficiency. (A) Mouse body weight relative to weight at the initiation of treatment (n = 7–15 mice/group). (B–O) Duodenal crypt regeneration was assessed at (B–H) 3 days post irradiation (DPI) and (I–O) 5 DPI by (B, C, I, and J) H&E staining, and (E, F, L, and M) EdU incorporation. (D and K) Villus height (n = 5–8 mice/group), (G and N) cellular proliferation, and (H and O) crypt regeneration were measured (n = 4–5 mice/group).

(P) Schematic of organoid formation assay to test stem cell activity in non-irradiated Villin-CreER^T2 mice. Duodenal crypts were isolated from TX- or VEH-treated mice and plated in Matrigel to form organoids.

(Q and R) Bright-field images of organoids 3 days post establishment from crypts isolated from (Q) VEH-treated or (R) TX-treated Villin-CreER^T2 mice.

(S) Organoid-forming efficiency was determined by counting organoid number and presented as percent of the number plated (n = 3 mice/group with three technical replicates per mouse).

Quantitative data are presented as means ± SEM (^∗p < 0.05, ^∗∗p < 0.01, ^#p < 0.0001 TX versus VEH by Student's t test). Scale bars, 100 μm (duodenum) and 250 μm (organoids). See also Figures S1 and S2.