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. 2019 Dec 27;9:20086. doi: 10.1038/s41598-019-56560-2

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© The Author(s) 2019

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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Calcium oxalate, monosodium urate and ATP induce IL-1β release in LPS-primed murine BMDCs, BMDMs and human PBMCs in vitro. (A,B) Murine bone marrow-derived dendritic cells (BMDCs), (C) murine bone marrow-derived macrophages and (D) human peripheral blood mononuclear cells were primed with LPS (100 ng/ml) for 3 hours before either calcium oxalate crystals (100 µg/ml) (OX), monosodium urate crystals (300 µg/ml) (MSU) or ATP (5 mM) or no further stimulus as control (CO) were added for an additional 6 hours. Culture supernatants (SN) were collected and concentrations of IL-1ß were measured using ELISA. (A,C,D) In addition, supernatants and whole cell lysates (LY) from stimulated murine BMDCs were analyzed by western blotting. (B) All three stimuli induced IL-1ß release in all three cell types. Data are presented as mean ± SD of a representative experiment of a total of nine each performed with triplicate biological samples. Statistical analysis was performed using two-way ANOVA and post hoc analysis. ****P < 0.0001; **P < 0.01; *P < 0.05; ns, not significant compared with control treatment (without LPS). To improve the clarity of the presented western blots, blots are displayed in a cropped version. Full-length gels are presented in Supplementary Fig. 1.