Fig. 1: Characterization of the effects of a mutation in mexZ for bacterial localization during colonization of Air Liquid Interface (ALI) airway infection models. | Nature Communications

Fig. 1: Characterization of the effects of a mutation in mexZ for bacterial localization during colonization of Air Liquid Interface (ALI) airway infection models.

From: Mutations in the efflux pump regulator MexZ shift tissue colonization by Pseudomonas aeruginosa to a state of antibiotic tolerance

Fig. 1

a Confocal images of the internal part of ALI cultures derived from primary cells of a person with Cystic Fibrosis after 14 h of infection with PAO1 or mexZ* P. aeruginosa in green (GFP), epithelial structure visualized by F-actin staining in red (Phalloidin) and nuclei in blue (To-pro), and their corresponding cross sections. Arrows in the cross section highlight the layer shown in the internal part image. Scale bar = 100 µm. The results shown were consistently obtained in three independent biological replicates of the experiment. b Confocal images of the internal part of fully differentiated BCi-NS1.1 cells ALI cultures after 14 h of infection with PAO1, mexZ* or mexZ*ΔlecA P. aeruginosa in green (GFP), epithelial structure visualized by F-actin staining in red (Phalloidin) and nuclei in blue (To-pro), and their corresponding cross sections. Arrows in the cross section highlight the layer shown in the internal part image. Scale bar = 100 µm. The results shown were consistently obtained in three independent biological replicates of the experiment. c Schematic representation of the ALI airway infection model set up. A pseudostratified layer of differentiated epithelial cells in contact with the air from its apical side and with medium from its basolateral side through the pore membrane of the transwell in which the human cells are located, is represented. When starting an infection to be monitored, P. aeruginosa cells (represented by green rods) are added to the apical side of the human cell culture. d Colony Forming Units (CFUs) of PAO1, mexZ* and mexZ*ΔlecA in the apical (red) and basolateral (blue) ALI compartments, and attached to the cell layer (green), after 14 h of infection in fully differentiated BCi-NS1.1 cells. Data are presented as mean values ± SEM of the results from three biological replicates with three technical replicates each. Statistical significance was determined by two-sided t-test assuming equal variances for CFU measurements and indicated as **** (p ≤ 0.00005): attached CFUs PAO1 vs mexZ* p = 0.00002; Attached CFUs mexZ* ΔlecA vs mexZ* p = 0.00005. Source data are provided as a Source Data file.

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