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. 2021 Jul 2;12:4105. doi: 10.1038/s41467-021-24352-w

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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. 6 — a Sorted WT bone marrow-derived macrophages (BMDMs) and FL3TL-pDCs were placed into the same wells for 16 h before addition of R848. TNF production in macrophages was measured by intracellular staining in macrophages at 4 h (n = 4 mice per group). b WT BMDMs were treated with recombinant IL-6 at various concentrations, and macrophage TNF or IL-12p40 production was measured by intracellular staining at 4 h. Percentage changes was normalized against WT samples without IL-6 (n = 3 mice per group). c, d Sorted WT FL3TL-pDCs were treated with R848 for 24 h, and their supernatants (pDC^sup) or rIL-6 (50 ng/ml) were added to Zc3h12c^+/+(blue)/Zc3h12c^−/− (red)BMDMs along with R848. TNF production in macrophages at 4 h (c) or 16 h (d) was measured by intracellular staining. Percentage changes was normalized against Zc3h12c^+/+ samples without pDC^sup or rIL-6 (n = 4 mice per group). e TNF production by Zc3h12c^+/+/Zc3h12c^−/− BMDMs with sorted WT BM pDC supernatants was measured by intracellular staining after R848-treatment at 4 h (n = 5 mice per group). f TNF production by Zc3h12c^+/+/Zc3h12c^−/− BMDMs with titrated concentrations of R848-treated sorted Zc3h12c^−/− FL3TL-pDC supernatants, with or without IL-6 neutralizing antibodies was measured by intracellular staining at 4 h (n = 3 mice for Zc3h12c^+/+; n = 4 mice for Zc3h12c^−/−). g IL-12p40 production by Zc3h12c^+/+/Zc3h12c^−/− BMDMs with pDC^sup or rIL-6 was measured by intracellular staining at 4 h (n = 5 mice for Zc3h12c^+/+; n = 4 mice for Zc3h12c^−/−). h IL-12p40 production by Zc3h12c^+/+/Zc3h12c^−/− BMDMs with titrated concentrations of R848-treated sorted Zc3h12c^−/− FL3TL-pDC supernatants, along with or without IL-6 neutralizing antibodies was measured by intracellular staining at 4 h (n = 4 mice per group). Data are representative of five (c, d), three (g), or two (a, b, e, f, h) independent experiments (mean ± S.D.; two-tailed Student’s t test). Source data are provided as a Source Data file.