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. 2019 Oct 29;9:15566. doi: 10.1038/s41598-019-52208-3

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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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Localization of osteopontin (OPN) in small airways. Immunoreactivity for OPN was visualized via reaction with 3,3′-diaminobenzidine (DAB), resulting in brownish staining. OPN was observed on the apical surface of the airway mucosa as well as in cells scattered within the epithelial cell layer of small airways in a never-smoker (A), smoker without chronic obstructive pulmonary disease (COPD) (B), and patients with COPD at various stages of the Global Initiative for Chronic Obstructive Lung Disease (GOLD) classification (C–E). Analysis of immunoreactivity within the epithelial cell layer was performed and groups were compared via one-way analysis of variance (ANOVA) with Dunnett’s post-hoc test. (F) *P < 0.05. SA = Small airway. Scale bars: A–D = 50 µm, E = 150 µm.