Gating strategy for the identification of L. pneumophila -containing neutrophils. Mice were infected with 2.5 × 106 CFU of L. pneumophila per mouse. Uninfected mice received PBS. Lungs were harvested at various times after infection and analysed for the presence of neutrophils that contained intracellular bacteria. A. Representative plots showing the presence of CD45+CD11b+Ly6G+ neutrophils in the lungs of infected mice. B. Representative plots showing the staining of CD45+CD11b+Ly6G+ pulmonary neutrophils from infected or uninfected mice with an anti-L. pneumophila antibody (Infected and Uninfected) or from infected mice stained with an isotype-matched antibody with an irrelevant specificity (Isotype).