Fig. 3: Cell cycle interference reduces the complexity of lumina.
a, A schematic showing the experimental design of the aphidicolin assay. Scale bar, 10 µm. b, Immunofluorescence images of DMSO- and aphidicolin-treated branching organoids expressing GFP–LifeAct at day 6. Scale bar, 20 µm. c, A 2D quantification (at mid-plane) of lumen number (left) and lumen occupancy (right) per organoid of DMSO- and aphidicolin-treated branching organoids at day 6. The data are represented as boxplots showing the median (centre line), the 25th and 75th percentiles (box edges) and whiskers extending to 1.5× the interquartile range. The data are pooled from three independent experiments (n = 17 DMSO; n = 34 aphidicolin). d, In silico simulations of organoids under τV = 40 and ξ = 0.12. Top: simulation run without stopping cell division. Bottom: a run in which cell division was stopped at 32 cells (red, cells; blue, lumen). e, A quantification of the relationship between the number of cells and mean organoid radius during the simulation (red, control with no stop in cell division; blue, cell division stopped at 32 cells). N = 50 simulations; the bold line represents the moving average ± standard deviation error bars. f, A quantification of lumen occupancy versus mean organoid radius during the simulation (red, control with no stop in cell division; blue, cell division stopped at 32 cells). N = 50 simulations; the bold line represents the moving average ± standard deviation error bars. g, A quantification of lumen number versus mean organoid radius during the simulation (red, control with no stop in cell division; blue, cell division stopped at 32 cells). N = 50 simulations; the bold line represents the moving average ± standard deviation error bars. h, The relationship between lumen occupancy (in two dimensions) and organoid radius of branching organoids treated with DMSO or aphidicolin (linear regression ± 95% confidence interval). The data are pooled from three independent experiments (n = 17 DMSO; n = 34 aphidicolin). i, The relationship between lumen number (in two dimensions) and organoid radius of branching organoids under DMSO and aphidicolin treatment (polynomial regression (order of 2) ± 95% confidence interval). The data are pooled from three independent experiments (n = 17 DMSO; n = 34 aphidicolin). The P values were determined by a two-sided Mann–Whitney test for c.
