Fig. 1

Co-culture establishment of 2D-airway organoids with P. aeruginosa PAO1. (a) Schematic representation of line establishment, 3D organoid expansion and 2D ALI differentiation. (b) HE staining of the major cell types present in ALI differentiated airway organoids, goblet cells stained by MUC5AC, ciliated cells by acetylated (Ac) tubulin and basal cells by TP63. Scale bar indicates 25 μm (c) Schematic representation of co-culture establishment of PAO1-GFP and differentiated airway organoids. (d) Z projections and (e) cross-section of confocal imaging of the co-culture after 14 h. Red: F-Actin; green: PAO1-GFP; blue: DAPI. Scale bar indicates 10 μm. (f) CFU assay of WT PAO1 bacteria and PAO1 ΔQS and ΔpqsA strains following co-culture with organoids and in liquid medium at time points 0 h and 14 h. The mean of the triplicates was plotted and error bars represent standard error of the mean (SEM). To determine statistical significance between the time points, log-transformed data was analyzed using two-way ANOVA, corrected for multiple comparisons using Sidak’s test (*P < 0.05; **P < 0.005; ****P < 0.0001).