Extended Data Fig. 3: Wnt ligands increase regenerative capacity of ISCs.
From: Notum produced by Paneth cells attenuates regeneration of aged intestinal epithelium
a, Distribution of organoid size on day 5 in ENR + 100 ng ml−1 Wnt3A ± 1μg ml−1 recombinant Notum (n = 50 organoids for Notum-treated (red), n = 38 organoids for untreated (black)). b, Area of colonies from sorted Lgr5hi stem cells from young and old mice (n = 3 mice per age group). Area quantified at day 7. c, Organoid-forming capacity of crypts from young and old mice treated with 100 ng ml−1 Wnt3A. Starting frequency was quantified on day 2 and is represented relative to untreated control (n = 10 mice per age group). d, Primary and secondary regenerative growth of young and old organoids treated with or without 100 ng ml−1 Wnt3A for the first 2 days of culture. Primary organoids were quantified at day 6 and secondary organoids two days after subculturing. Data are represented relative to untreated control (n = 9 mice per age group). e, Organoid-forming capacity of isolated crypts from young mice treated with or without 1 μg ml−1 recombinant Notum (n = 3 mice). f, Primary regenerative growth of organoids from young mice treated with or without 1 μg ml−1 recombinant Notum, quantified on day 6 (n = 3 mice). g, Secondary regenerative growth of organoids from young mice treated with or without 1 μg ml−1 recombinant Notum, quantified on day 2 after subculture (n = 3 mice). h, Organoid-forming capacity of isolated crypts at day 2 from young mice treated with Porcupine inhibitor IWP-237 (n = 3 mice). i, Primary regenerative growth of organoids treated with IWP-2 for the first two days of culture. Organoids were quantified on day 6 (n = 3 mice). j, Flow cytometry analysis of cellular frequencies from Lgr5–eGFP organoids two days after treatment with IWP-2 (n = 4 mice). Y, mice between 3 and 9 months of age; O, mice over 24 months of age in all experiments. In box plots, unless otherwise indicated, the line represents median, the box shows interquartile range and whiskers represent the range. All other data are mean ± s.d.; two-tailed unpaired Student’s t-test; exact P values shown in corresponding panels.
