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. 2020 Nov 5;10:19091. doi: 10.1038/s41598-020-76251-7

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Mechanistic study of the anti-inflammasome property of riboflavin. (A) LPS-primed BMDMs were treated with rotenone (Rot) with/without riboflavin (B2). The maturation and secretion of IL-1β and Casp1 were analyzed by immunoblotting and the secretion of IL-1β was analyzed by ELISA. Diphenyleneiodonium chloride (DPI) was used as a ROS scavenger. (B) LPS-primed BMDMs were treated with ATP in the presence of B2. The cytosolic release of mitochondrial DNA (mtDNA, cytochrome c oxygenase 1/18S rDNA) was measured by quantitative real-time PCR. (C) The activity of human recombinant caspase-1 (hrCasp1) in the presence of B2 was measured using a commercial kit, which is a modified luciferase assay. Briefly, hrCasp1 catalyzes Z-WEHD-aminoluciferin to aminoluciferin, a substrate of luciferase, resulting in the generation of light, which represents the activity of Casp1. Ac-YVAD-CHO (YVAD, the kit supplied) was utilized as a caspase-1 inhibitor. All immunoblot data shown are representative of at least two independent experiments. The bar graph presents the mean ± SD with at least two independent experiments.