Extended Data Fig. 7: Theoretical controls and sensitivity analysis.
a–c, Comparison between experimental data (dots and error bars) and theoretical predictions (thick lines) for the time evolution of the clonal rootedness (a), average clone size (b) and clonal persistence (c), for both KRT19 (cyan) and the LGR5 (green) tracings from E16.5. Here, the theoretical prediction corresponds to the case of villi fission, following exactly the same model in Extended Data Fig. 6, but stopping at P1 instead of at P5. This shows that fetal fission is enough to explain the bulk of the equipotency between LGR5 and KRT19 clones. d–f, Comparison between experimental data (dots and error bars) and theoretical predictions (thick lines) for the time evolution of the clonal rootedness (d), average clone size (e) and clonal persistence (f), for both KRT19 (cyan) and LGR5 (green) tracings from E16.5. Here, the theoretical prediction corresponds to the case of no new villi formation occurring, showing a very poor fit for the clonal persistence and size to the data. g–i, Comparison between experimental data (dots and error bars) and theoretical predictions (thick lines) for the time evolution of the clonal rootedness (g), average clone size (h) and clonal persistence (i), for both KRT19 (cyan) and LGR5 (green) tracings from E16.5. Here, the theoretical prediction corresponds to the case of villi fission occurring as in the model shown in Fig. 3, but without a shift of cells upon villi fission (see schematics in Extended Data Fig. 5), showing a very poor fit for the clonal persistence to the data. j, Cartoon illustrating that the current villification model suggests that villi emerge from the intervillus region. k–m, Comparison between experimental data (dots and error bars) and theoretical predictions (thick lines) for the time evolution of the clonal rootedness (k), average clone size (l) and clonal persistence (m), for both KRT19 (cyan) and LGR5 (green) tracings from E16.5. Here, the theoretical prediction corresponds to the case of villi formation occurring only from existing crypts, showing a very poor fit for the clonal persistence to the data. n, Volume of single villus cells and intervillus–crypts cells. Total E-cadherin volume of villi and intervilli–crypts, subsequently divided by the number of DAPI+ nuclei to estimate single-cell volume. E16.5 villi n = 8, E16.5 intervilli n = 10, P0 villi n = 6, P0 intervilli n = 3, P5 villi n = 11, P5 intervilli n = 4, P11 villi n = 4, P11 intervilli n = 4, adult villi n = 7, adult intervilli n = 4 independent images were analysed. o, Sensitivity analysis on the influence of differential LGR5− and LGR5+ cellular volume on the model prediction—considering either that all cells have the same volume (continuous lines and a–n) or the differential volume measured at E16.5 (see Supplementary Theory Note, dashed lines). For a–i, k–m: P0 n = 3, P5 n = 3, P11 n = 3, adult n = 6 independent samples analysed for LGR5; P0 n = 1, P5 n = 3, P11 n = 6, adult n = 3 independent samples analysed for KRT19. Data are mean ± s.e.m.
