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. 2021 Dec 24;11:24432. doi: 10.1038/s41598-021-04133-7

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© The Author(s) 2021

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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Sars-Cov-2 E-protein suppresses the ER stress response, inflammasome priming and activation. (A) Quantitative real-time PCR analysis of ER stress genes in bone marrow derived macrophages (BMDMs). Data are presented as mean ± SD, which were analyzed by unpaired t-test. ***P ≤ 0.001, **P ≤ 0.01, *P ≤ 0.05. (B) Quantitative real-time PCR analysis of inflammasome genes in BMDMs. Data are presented as mean ± SD, which were analyzed by unpaired t-test. ***P ≤ 0.001, **P ≤ 0.01, *P ≤ 0.05. (C) Western blot analysis of inflammasome proteins in cellular lysates of Wild type (WT) and Nlrp3^−/− BMDMs, transduced with control and E-protein lentiviruses for 72 h and were primed with 20 ng/ml LPS for 4 h. Blots were cut to probe with NLRP3 (from ~ 80 kDa to top) or pro-caspase-1, GSDMD, pro-IL-1β and Actin (from ~ 80 kDa to bottom). (D) Western blot analysis of inflammasome proteins in cellular lysates of WT and Nlrp3^−/− BMDMs, transduced with control and E-protein lentiviruses for 72 h and were primed with 20 ng/ml LPS for 3 h and treated with Nigericin for 1 h. (E) Quantification of IL-1β secretion into cell media in WT and Nlrp3^−/− BMDMs. BMDMs were transduced with control and E-protein lentiviruses for 72 h and then primed with 20 ng/ml LPS for 3 h and treated with Nigericin for 1 h. IL-1β secretion was assessed via IL-1β ELISA kit. Data are presented as mean ± SD, which were analyzed by one-way ANOVA coupled with Tukey’s test for multiple comparisons. ***P ≤ 0.001, **P ≤ 0.01, *P ≤ 0.05. (F) Quantification of IL-18 secretion into cell media in WT and Nlrp3^−/− BMDMs. BMDMs were transduced with control and E-protein lentiviruses for 72 h and then primed with 20 ng/ml LPS for 3 h and treated with Nigericin for 1 h. IL-18 secretion was assessed via IL-18 ELISA kit. Data are presented as mean ± SD, which were analyzed by one-way ANOVA coupled with Tukey’s test for multiple comparisons. ***P ≤ 0.001, **P ≤ 0.01, *P ≤ 0.05.