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. 2019 Nov 15;9:16893. doi: 10.1038/s41598-019-53231-0

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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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Heme impairs IL-36-mediated cytokine production in human patient FLS. FLS from RA patients treated with heme (A) or two ratios of heme-IL-36α (B) for 24 h. Relative mRNA expression of IL-6 and IL-8, normalized to GAPDH, analyzed by quantitative real time PCR (qRT-PCR; top), and cytokine concentrations in the cell culture supernatant measured by ELISA (bottom). Values are means + SEM. One representative experiment (n = 6) of two independent experiments is shown. Statistical analysis was performed using Mann-Whitney Test Bonferroni corrected with *p < 0.05 and **p < 0.01. (C) FLS from RA patients treated with varying ratios of IL-36α or IL-36α mutants to heme for 24 h (light and dark grey bars). Relative mRNA expression of IL-6, normalized to GAPDH, and analyzed by qRT-PCR. Values are means ± SEM of one experiment (n = 3). Statistical analysis was performed using One-Way Anova Test Dunnet corrected, with *p < 0.05, **p < 0.01, and ***p < 0.0001 for comparisons with recombinant trIL-36α.