Extended Data Fig. 1: Development of in vitro lung tissues.

a. Schematics of the cell culture process to develop fully differentiated (+21 days post airlift) airway epithelium on Transwell inserts and subsequent apical infection. b, c. Development of the lung tissue post airlift (day 0 to 60) visualized by histology (b) or ICC (c). d, e. Quantification of the thickness (d) and total cell numbers (e) of tissues at day 0, 30 or 60 post airlift, inferred from ICC images. For (d) and (e): Statistical significance was calculated using Kruskal–Wallis H-test with Dunnett’s multiple comparison test. ns; not significant. For panels a-c: n = 3 independent experiments (same donor). For d and e.: n = 15 3D images (30 regions/images for d) examined over 3 independent experiments. Data are presented as mean values +/− SD. f, g. Comparison of airway epithelium on Transwell inserts from 3 donors visualized by histology (f) or ICC (g). n = 3 independent experiments (different donors). Histology: hematoxylin and eosin staining. ICC staining: nuclei: DAPI, mucus: Muc5AC, actin: phalloidin-647, cilia: acetylated-α-tubulin.