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. 2018 Jan 10;8:256. doi: 10.1038/s41598-017-18433-4

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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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IL-34 macrophages have a different polarization potential as compared to CSF-1-macrophages. (a) Human monocytes were differentiated during 7 days with 100 ng/mL CSF-1 or 100 ng/mL IL-34 and then polarized into M1-macrophages (LPS + IFNγ) or M2-macrophages (IL-4) for 2 days. Macrophage polarization was evaluated by 2-color flow cytometric analysis. (b) Functional assay of monocytes exposed for 2 or 5 days to 100 ng/mL CSF-1 or 100 ng/mL IL-34. The results are expressed as MFI and represent the mean ± SD of 3 independent experiments performed in duplicate. n.s. denotes not statistically significant according to a paired student t test. (c) Functional assay of monocytes exposed for 7 days with 100 ng/mL CSF-1 or 100 ng/mL IL-34 and then polarized into M2-macrophages (IL-4) for 2 days. The results are expressed as the fold induction compared to CSF-1 macrophages and represent the mean of 3 independent experiments performed in duplicate. *P < 0.05 according to a paired student t test (versus CSF-1-macrophages). (d,e) Human monocytes were differentiated during 7 days with 100 ng/mL CSF-1 or 100 ng/mL IL-34 and then polarized into M1-macrophages (LPS + IFNγ) or M2-macrophages (IL-4) for 24 hours. The expression of the indicated mRNA was analyzed by qPCR (mean ± SEM of 5 independent experiments). *P < 0.05, ***P < 0.001 according to a paired student t test (versus CSF-1 macrophages). The production of IL-10 and CCL17 was analyzed using Multi-Analyte ELISArray kit as described in Material and Methods section. The results are expressed as ng/mL per million of cells and represent the mean ± SD of 2 independent experiments performed in duplicate.