Fig. 5.
Adenoma-derived organoids exhibit increased dependence on EGFR signalling. a, b Organoids were generated from the normal epithelium or adenomas of the ApcΔ716 mouse small intestine, and infected with lentiviruses expressing a FRET biosensor for ERK activity (EKAREV-NLS). a Representative images of ERK activity (FRET/CFP ratio) in organoids derived from the normal epithelium and adenomas. b Bee swarm plots of ERK activity in the normal epithelium-derived and adenoma-derived organoids (Normal: n = 61, Adenoma: n = 99 cells, from more than three organoids). c–f Adenoma-derived organoids expressing EKAREV-NLS were treated with 1 μM of an EGFR inhibitor (EGFRi), PD153035, and/or 10 μM of an ErbB2 inhibitor (ErbB2i), CP-724714. c Representative images of adenoma-derived organoids before and after treatment with EGFRi or ErbB2i. d Bee swarm plots of ERK activity before and after inhibitor treatment (EGFRi: n = 120, ErbB2i: n = 119, EGFRi + ErbB2i: n = 109 cells, pooled from three organoids). e, f Quantification of ERK activity pulses in adenoma-derived organoids. Organoids were treated with EGFR and/or ErbB2 inhibitors, and then imaged for 90 min. ERK activity data from each cell were processed as described for Fig. 4e (−/−: n = 40, EGFRi/−: n = 45, −/ErbB2i: n = 32, EGFRi/ErbB2i: n = 29 cells). Frequencies of the pulse-like ERK activity (e) and the proportion of cells with pulse-like ERK activity (ERK-pulse+) (f) under each condition. Scale bars, 50 µm. Red lines represent mean. Error bars represent s.e.m. Mann–Whitney U-test (b, d) and Steel–Dwass test (e) were used for comparison. *P < 0.05, **P < 0.001, ***P < 0.0001, n.s., not significant