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. 2017 Aug 7;7:7471. doi: 10.1038/s41598-017-08264-8

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© The Author(s) 2017

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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Decreased numbers but M2 polarization of alveolar macrophages in Abt mice challenged with B16/F10 melanoma. The mice were given antibiotics for five weeks and then challenged with B16/F10 cells (1 × 10⁵ cells/mouse, i.v.). On day 17 after the B16/F10 challenge, the lung MNCs were isolated. (a) Frequency and number of alveolar macrophages (F4/80^hi CD11c^hi) in the lung MNCs from mice challenged with B16/F10 cells (n = 5). (b) Purified alveolar macrophages (F4/80^hi CD11c^hi) were analyzed by GeneChip. The pie chart showed the distribution of DEGs in Abt group compared with the control. Unknown and duplicated genes were filtered for the analysis (2 samples/group, 15 mice/sample). (c) Purified alveolar macrophages (F4/80^hi CD11c^hi) (3 × 10⁵ cells/well) were stimulated for 48 h in the DMEM containing 10% FBS with the stimulation of 1 µg/ml LPS, 100 µg/ml PolyI:C, 1 µg/ml flagellin or 20 ng/ml PMA, and then culture supernatants were analyzed for the cytokine expression by ELISA (Arg1 and CCL24) and CBA (IL-13, IL-6, IL-10 and IL-1β) (n = 3). The data are shown as the mean ± SEM. *p < 0.05 compared with the control group.