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. 2020 Aug 6;10:13212. doi: 10.1038/s41598-020-69984-y

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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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BCG infection induced the elevation in the counts of IL-35-producing B cells in the blood and in each immune tissue. PBMCs and mononuclear cells from the lung, spleen, and bone marrow were extracted from BCG-infected mice (n = 6). Then, the cells were assessed for the frequencies of IL-35-producing B cells by flow cytometry. The gating strategy is shown in Figure S2. (A, B, E, F) shows the representative histograms for flow cytometry analysis of the expression of IL-35 in B cells in PBMCs (A) and B cells in mononuclear cells from lung (B), spleen (E), and bone marrow (F) in BCG-infected mice and controls for 4 weeks. (C, D, G, H) shows the graph data showing the mean frequencies of IL-35-producing cells in B cells in PBMCs (C) and in mononuclear cells from the lung (D), spleen (G), and bone marrow (H) in BCG-infected mice (n = 10) for 2, 4, and 8 weeks and controls (n = 6). The P value is shown in each column. *P < 0.05; **P < 0.01; ***P < 0.001.