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. 2020 Jul 23;6:63. doi: 10.1038/s41420-020-00300-3

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© The Author(s) 2020

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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Fig. 5 — a The self-renewing macrophages were cultured for 5 days with rhM-CSF (upper) alone or rmGM-CSF plus rmIL-4 (lower). b The self-renewing macrophages were cultured as in a, and analyzed for the expression of CD80, CD11c, or MHC-II by flow cytometry. MFI mean fluorescence intensity. c The self-renewing macrophages were cultured for 7 days with rhM-CSF (upper) alone or rhM-CSF plus rhRANKL (lower), and subjected to tartrate-resistant acid phosphatase (TRAP) staining. d The self-renewing macrophages were cultured as in c for 5, 6, or 7 days. Their TRAP activity was also quantified using the TRAP solution kit, and calculated as nmoles of p-NP per ml (mean ± SD, n = 3). e The self-renewing macrophages were cultured as in c for 3 days, and analyzed for the expression of the indicated genes by qRT-PCR.