Extended Data Fig. 2: Cellular composition and functional properties of the bronchial lung epithelium model.

a, b. Visualization (a) and quantification (b) of the major cell types present in the bronchial epithelium by ICC or FACS, respectively. n = 2 independent experiments (same donor). c. Measurements of cilia beating frequencies by microscopy at day 30 or 60 post airlift. n = 36 inserts over 3 independent experiments (same donor); Box plot: center line, median; box limits, upper and lower quartiles; whiskers, min-max. d, e. Assessment of epithelial barrier integrity (TEER) of lung epithelium at day 0, 30 or 60 post airlift (d, n = 3 independent experiments, same donor) and epithelia from 3 different donors (e, n = 5 independent experiments). For (c), (d) and (e): statistical significance was calculated using Kruskal–Wallis H-test with Dunnett’s multiple comparison test. ns; not significant. f. Visualization of tight junctions by ICC of lung epithelium at day 0 or 30 post airlift. g. Assessment of epithelial barrier integrity (Lucifer yellow permeability) of lung epithelium from 3 different donors (n = 6 inserts over 3 independent experiments). h. Wound healing after physical tissue damage (indicated by the white arrow) of the lung epithelium was followed by time-lapse live imaging for 36 hours (see: Supplementary Information Movie 2). For b, c and f: data are presented as mean values +/− SD. For d, e and g: data are presented as mean values +/− SD. Staining: nuclei: DAPI, cycling basal cells: mucus: Muc5AC, cycling basal cells: α-Ki67, basal cells: α-p63, club cells: α-SCG1A1, cilia: acetylated-α-tubulin, actin: phalloidin-647, tight junctions: ZO-1 Alexa Fluor 555, membrane: CellMask.