A Model of Intracellular Persistence of Pseudomonas aeruginosa in Airway Epithelial Cells

Figure 2

Analysis of P.a.-infected AEC by flow cytometry. (a) Gating strategy based on mCherry fluorescence intensity and forward scatter area (FSC-A) to detect mCherry(+)-infected AEC, with exclusion of mCherry(-) noninfected AEC using control samples infected with nonfluorescent PAO1. (b) mCherry fluorescence histograms of the AEC populations at different time points after infection with nonfluorescent PAO1 (whole population, grey) or mCherry PAO1 (mCherry(+) population only, colored) as indicated. Quantification of the proportion of (c) mCherry(+) AEC among all live AEC or (d) dead (FVD+) AEC among all AEC. (e) mCherry median fluorescence intensity (MFI) of the mCherry(+) AEC population at different time points after infection. The mCherry MFI of cells infected with nonfluorescent PAO1 was subtracted from the MFI of mCherry(+) cells to account for day-to-day variability in fluorescence. For all experiments, BEAS-2B cells were infected at MOI 1, except for uninfected (UI) controls. ≥20,000 cells were analyzed in each sample. For (a) and (b), representative scatter plots and histograms are shown. For (c), (d), and (e), results shown are from biological triplicates from one representative experiment of 3 independent ones (data from all 3 independent experiments are shown in Figure S2). ; ; .

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