Fig. 3: Tsc1 deletion leads to defects in propagation of organoids and ISC/progenitor cells.

a, b Eight-month-old Lgr5-GFP-CreERT;Tsc1^f/f mice showed decreases in the number of IECs (a) and the number of proliferating TA cells (b) (based on Fig. 2b). Data are expressed as mean ± SEM. N = 4 mice per group. **P < 0.01 (determined using Student’s t test). c Eight-month-old Lgr5-GFP-CreERT;Tsc1^f/f mice showed no change in the villus-to-crypt ratios. N = 4. d Representative images showing that Tsc1^−/− crypts (isolated from Villin-Cre;Tsc1^f/f mice, since Lgr5-GFP-CreERT can label ~65% of the crypts) displayed increases in the size of the minigut organoids and the number of crypts, whereas serial passaging of the organoids revealed that Tsc1-deficient organoids quickly lost their ability to propagate. Right panels: quantification of the data (mean ± SEM). N = 3 mice per group. *P < 0.05, **P < 0.01 (determined using Student’s t test). e Eight-month-old Lgr5-GFP-CreERT;Tsc1^f/f mice showed decreases in the numbers of ISCs and proliferating ISCs. Intestine sections were immunostained for PCNA, and cells positive for PCNA and GFP were counted and normalized to the total numbers of Lgr5⁺ ISCs. Bottom panels: quantitation data (mean ± SEM). N = 5 mice per group. **P < 0.01 (determined using Student’s t test). f The numbers of ISCs and proliferating ISCs were decreased in naturally aged 17.5-month-old mice. Right panels: quantitation data (mean ± SEM). N = 3 mice per group. *P < 0.05, **P < 0.01 (determined using Student’s t test).