Extended Data Fig. 9: Perturbation of FAK and YAP after chemotherapy.

a. (left) Time-point live imaging of LGR5-tdTomato/p27-mVenus CCO32 organoids following treatment with CPT (10 nM, 24 hr) treatment alone or in combination with FAKi (10 μM, continuous) (left). LGR5-tdTomato; red and p27-mVenus; green. The experimental timeline is shown on the top. The transition of the percentage of LGR5⁺p27⁺ in all cells (mean ± s.e.m.) in CCO32 organoids following the indicated treatments (right). *p < 0.05, linear model, followed by Tukey adjustment (control vs CPT + FAKi; p = 0.0020, CPT vs CPT + FAKi; p = 0.013, FAKi vs CPT + FAKi; p = 0.013). b. Representative YAP staining (magenta) and p27-mVenus (green) expressions in sorted LGR5⁺p27⁺ cells at the indicated time points after sorting (left). Cells were treated with CPT (10 nM, 24 hr) before sorting. Representative confocal images of short-cultured LGR5⁺p27⁺ CCO32 cells with EGFR inhibition (1 μM) at the indicated time points after single-cell sorting (right). LGR5 (red), p27 (green), YAP staining (magenta). c. The cell counts in cells/clusters derived from sorted single LGR5⁺ p27⁺ cells at the indicated time points after sorting. The organoids were treated with CPT (10 nM, 24 hr) before sorting. Each cell is coloured based on the YAP and p27 status. d. The distribution of the cell counts in cells/clusters derived from sorted single LGR5⁺ p27⁺ cells on day 3 after sorting. The organoids were treated with CPT (10 nM, 24 hr) before sorting, and with or without 20 μM TEADi after sorting. Red points and lines indicate mean ± sd. p = 1.7 x 10^–6, two-sided Wilcoxon rank sum test. Scale bars: 10 μm (b), 100 μm (a)

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