Fig. 2.
Stem cell replacement rate is accelerated after LGK974 treatment. Mice were induced with 0.15 mg tamoxifen to induce tdTomfl recombination in few Lgr5-CreER-EGFP+ cells. a Mosaic expression of the Lgr5-eGFP (green) and the recombined tdTomfl+ cells (red). Nuclei were stained with DAPI (blue). Representative pictures at day 10, note the loss of GFP expression and clones are fully labelled by tdTom+ cells in Porcupine inhibitor-treated mice (LGK974). Scale bar = 100 um. b Clone size was counted in 'eighths', at time indicated after induction. At least 200 clones per mouse were counted, vehicle (VEH) N = 3, 3, 4, 4, 3, 3 and LGK974 (LGK) N = 2, 4, 3, 5, 3, 3 for each timepoint, respectively. Heatmap shows all counted clones per timepoint/group. Note that LGK974 has an increased clone size at day 4 and the mean clone size from day 7 is almost at its maximum. Graph shows the mean clone size (c) or the number of fully fixed crypts (d) at different time points as shown in b. Error bars, s.e.m. e Number of tdTom+ clones per field. The number of crypts with at least one tdTom+ cell were counted per field, ≥19 images per mouse, error bars = s.e.m. Note the similar number of clones at day 4 but the greater reduction in clones after LGK974 treatment at day 10. Vehicle (VEH) N = 3, 3, 4, 4, 3, 3 and LGK974 (LGK) N = 2, 4, 3, 5, 3, 3 for each timepoint respectively