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. 2019 Aug 22;9:12277. doi: 10.1038/s41598-019-48794-x

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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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Celastrol inhibits ATP-induced activation of NLRP3 inflammasome in primary macrophages. (A) Chemical structure of celastrol. (B–E) Primary mouse macrophages were primed with LPS (100 ng/ml) for 4 hr. The cells were treated with celastrol for 1 hr and then stimulated with ATP (5 mM) for (B,D,E) 1 hr or (C) 2 hr. (B) The cell culture supernatants and cell lysates were immunoblotted for pro-caspase-1, caspase-1(p10), pro-IL-1β, and IL-1β. (C) The cell culture supernatants were analyzed for secreted IL-1β using ELISA. The values represent the means ± SEM (n = 3). *Significantly different from ATP alone, p < 0.05. (D) Cell lysates were processed for immunoblotting of ASC. (E) The cells were stained for ASC (green). The nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue). The arrows indicate ASC speckles. The data are representative of three independent experiments. For immunoblotting results, the cropped blots from full length gels were presented. DIC, differential interference contrast.