A, B. Number of bromodeoxyuridine (BrdU)+ crypt base columnar cells after a 4-hour pulse in MHC-IIL/L; Villin-CreERT2 (vil-iKO) mice after tamoxifen administration (WT: n=3, vil-iKO: n=4, mean ± s.d). Representative images of BrdU immunostain in proximal small intestinal crypts (B).
C-E Organoid-initiating capacity of WT and vil-iKO crypts (C, n=5, mean ± s.d). Number of secondary organoids per dissociated crypt-derived primary organoid (D, n=5, mean ± s.d). Representative images of day-5 WT and vil-iKO primary organoids (E).
F. Relative expression of MHC-II in dissociated WT and vil-iKO primary organoids at day 5 (n=5, mean ± s.d).
G, H. Organoid-initiating capacity of ISCs from WT and MHC-IIL/L; Lgr5-EGFP-IRES-CreERT2 (Lgr5-iKO) mice with and without Paneth cells (P) from WT mice (n=5, mean ± s.d). Representative images of organoids derived from WT and Lgr5-iKO ISCs co-cultured with WT Paneth cells five days after seeding (H).
n.s.: not significant, ***P < 0.001 (Student’s t-tests). Scale bars, 100 μm (E, H) and 20 μm (B).
See also Figure S2