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. 2018 Apr 30;8:6777. doi: 10.1038/s41598-018-25167-4

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© The Author(s) 2018

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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Migrated neutrophils show increased cell surface expression of CD11B and MPO compared to non-migrated neutrophils. Neutrophils that migrated (M) to the apical compartment through RSV infected epithelial cells infected for (A) 24 hours or (B) 72 hours showed higher cell surface expression of i) CD11B and ii) MPO compared to those non-migrated (NM) neutrophils that remain on the basolateral side. The percentage of migrated and non-migrated neutrophils expressing CD11B and MPO was calculated by staining for cell surface expression of CD11B (PE) and MPO (APC) were determined by flow cytometry. Neutrophils were gated on initially using a PE positive gate. Using this population the geometric mean fluorescence intensity of PE and APC fluorescence was calculated. Bars show mean ± SEM of n = 4 biological repeats. Statistical significance is shown. For all other comparisons P > 0.05.