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. 2019 Oct 31;9:15703. doi: 10.1038/s41598-019-51991-3

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Coinfection with H. polygyrus increases neutrophil transepithelial migration into the P. aeruginosa infected airspace of the lung. FACS analysis using Ly6G and CD11b markers to isolate the neutrophil population in the (a) BAL, (b) lung tissue and (c) peripheral blood (PB), data are shown as mean ± SD, and are representative of three independent experiments. *P < 0.05, **P < 0.01, ****P < 0.0001 between the indicated conditions. (d) The left lung of the mice was stained with H&E, the black bar indicates 20 μm. (e) The infiltrating cells in the bronchi, small airways and parenchyma of H&E stained lung sections were counted using ImageJ software. (f) Lung sections were stained with FITC-labeled anti-mouse Ly6G. Sections were analyzed by confocal microscopy. (g) The fluorescence density of lung sections was enumerated by the ImageJ software (Adjusted Integrated density = Integrated Density-Area*Mean density of the background).