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. 2021 Oct 20;12(11):971. doi: 10.1038/s41419-021-04275-6

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© The Author(s) 2021

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. 8 — A The expression of B7–H3 mRNA was analyzed using RT-qPCR on human HC PBMCs and RA and AS synovial fluid macrophages. Monocytes were purified from PBMCs (n = 7), and macrophages were derived from RA synovial-fluid mononuclear cells (n = 9) and AS synovial-fluid mononuclear cells (n = 6). The results are shown as mean ± S.E.M. B The expression of B7–H3 protein between HC PBMCs and RA synovial-fluid macrophages (n = 3) was assessed by Western blot. C M-CSF (40 ng/ml) was added to RA synovial-fluid macrophages for three days. OCPs that were transiently transfected with B7–H3-specific siRNA or control siRNA (40 nM) were induced to differentiate using M-CSF (40 ng/ml) and RANKL (80 ng/ml) for seven days. TRAP staining was performed and the number of TRAP-positive multinucleated cells per well were counted as osteoclasts (scale bar, 200 μm). D The expression of the mature osteoclast markers, CTSK, TRAP, DC-STAMP, and ITGB3, and B7–H3, was analyzed by RT-qPCR. E, F The expression of B7–H3, NFATc1, IDO, phospho-STAT1 (PY701), and STAT1 protein was evaluated using Western blot. G The expression of IFN-inducible genes was analyzed using RT-qPCR at day 0. H HC PBMCs and RA synovial-fluid macrophages were treated with recombinant IFN-β (1 ng/ml) for one or two days. Whole-cell lysates were immunoblotted with B7–H3, IDO, phospho-STAT1 (PY701), and STAT1 antibodies. (A, D, G) The mRNA levels were normalized relative to GAPDH expression.