Air-liquid interface cultured differentiated human bronchial epithelial cell were performed (see Supplementary Material).19 (link) Calu-3 (Cat#CL-0054) cells were purchased from Procell. The human bronchial epithelial cell lines BEAS-2B and 16HBE are challenging to culture in the air-liquid interface cultured; whereas, the Calu-3 cell line can be cultured in the air-liquid interface for >14 days.20 (link)
P. aeruginosa PAO1 strain (ATCC) was resuscitated and incubated overnight in a shaker, and the logarithmic growth phase was diluted to different colony forming unit (CFU)s with LB broth and added to the upper chamber of the transwell.