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. 2017 Jun 14;7:3491. doi: 10.1038/s41598-017-03675-z

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© The Author(s) 2017

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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Andrographolide (ANDRO) affects inflammasome assembly in hepatic cells. Fat-laden hepatocytes were treated with LPS for a short period of time (3 h) and then were treated with ANDRO for 1 h. Total proteins were isolated and immunoprecipitated with an antibody against the inflammasome protein Apoptosis-Associated Speck-Like Protein Containing CARD (ASC). (A) The immunoprecipitated extract was separated by SDS-PAGE and then blotted and revealed with antibodies anti-Nucleotide-binding oligomerization domain leucine-rich-repeat containing receptor 3 (NLRP3) and Caspase-1 (CASP1) to assess inflammasome assembly (IP:ASC, left panel). CASP1 and NLRP3 immunoprecipitated with ASC in LPS-treated cells (IP:ASC, left panel, lane2) confirming LPS-induced inflammasome assembly. ANDRO treatment interfered with this process (IP:ASC, left panel, lane4). Before the immunoprecipitation assay, the presence of the indicated proteins was determined by western blot of total extracted proteins (Input control blot, right panel). (B) Anti-ASC immunoprecipitated proteins (CASP1 and NLRP3) were quantified and plotted over input protein levels.