Fig. 3: Neutrophil maturity diminishes over time in response to CSOM infection.

Mice underwent trans-tympanic occlusion of the Eustachian tube 1 day prior to infection with PAO1. Neutrophils from middle ear effusion of P. aeruginosa infected CSOM mice were assessed by flow cytometry 1, 14, and 28 days post infection. a A timeline for the treatment and analysis steps used in this model. b Violin plot of Ly6G-PE fluorophore depicts a reduction in Ly6G in chronically infected CSOM mice, indicative of a reduction in neutrophil maturity. A representative plot from effusion of mice at 14 d.p.i. is shown. c Manual flow cytometry gating strategy for Ly6G^int (immature)/Ly6G^hi (mature). Representative plots for uninfected control (L) and CSOM (R) are shown at 14 d.p.i. d Neutrophil subpopulations were quantified by the gating strategy in (c) at 1, 14, and 28 d.p.i. Average of 3–5 biological replicates are shown. (*p < 0.05 by unpaired t-test; error bars show SD).