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. 2019 Apr 8;9:5788. doi: 10.1038/s41598-019-42298-4

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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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EEA1 knock down results in a decrease release of IL-1β from macrophages without affecting pyroptosis. (A) Immunoblot analysis of EEA1, NLRP3, ASC, caspase-1 and IL-1β on cell lysate and cell-free supernatant from BMDMs transduced with non-target control (Non-target Lv) or EEA1 specific shRNA lentiviral particles (shRNA EEA1 Lv), primed with LPS and activated with ATP as indicated. This figure is representative of three independent experiments. (B) Densitometry quantification of EEA1 immunoblots shown in A. Relative EEA1 expression is normalized to β-actin expression. (C,D) ELISA for released IL-1β (C) or LDH (D) from BMDMs treated as in A. Nitrocellulose membranes were cropped after protein blotting and then inmunostained separately for each protein. Complete blots for IL-1β ans Caspase-1 are showed as supplementary. Data are presented as average ± sem from n = 3 independent experiments. ns, not significant (p > 0.05); ***p < 0.0001; *p ≤ 0.05.