Mice with LysMCre-mediated deletion of NOX2 exhibit an inflammatory lung phenotype. (A) BAL leukocyte count, flow-cytometric analysis of the percentage of polymorphonuclear leukocytes (PMNs), and total PMNs in 3 mL BAL from 12-week-old Ncf2^fl/fl and Ncf2^LysMCre mice (n ≥ 7). (B) ELISA analysis of BAL samples from 12-week-old mice (n ≥ 6). (C) Flow cytometric analysis of CD11c⁺CD11b^high macrophages from the CD45⁺Siglec F⁺ CD11c⁺ gate and the percentage that are CD11b^high in BAL samples of 12-week-old mice (n ≥ 8). (D) Lung sections from12-week-old Ncf2^fl/fl and Ncf2^LysMcre mice stained with hematoxylin-eosin showing intracellular (arrowhead) and extracellular (arrow) eosinophilic crystals. Bars represent 250 μm. Representative images from more than 6 samples per genotype. (E) tSNE plot from CyTOF data of CD45⁺CD11c⁺Siglec F⁺ AMs from lung tissue harvested from 12-week-old WT, Cybb^KO and Ncf2^LysMcre mice showing populations expressing CD11c, Siglec F, and CD11b. Representative plots from ≥4 samples in each group. Bar graph data are expressed as the mean ± standard error of the mean. The Student t test was performed for samples distributed in 2 groups. Data represented the results of at least 2 independent experiments. *P < .05; **P < .01; ***P < .001. CyTOF, cytometry by time of flight; t-SNE, t-distributed stochastic neighbor embedding.