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. 2020 Nov 6;10:19272. doi: 10.1038/s41598-020-76285-x

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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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Knockdown of chop alleviates immunoparalysis in CLP mice. (a) Adenoviruses expressing shRNAs specifically targeting chop were intravenously injected into C57BL6 mice through the tail vein. After 7 days, CLP was performed. Effects of chop-knockdown on the mortality of CLP mice was detected (n = 20), Kaplan–Meier curves and the log-rank test were used to compare mortality rates, *p < 0.05. (b) The effects of chop-knockdown on the bacterial clearance in blood, peritoneum lavage and spleen of CLP mice were examined at the indicated times after CLP (n = 6). (c) The effects of chop-knockdown on the production of IL-1β (c1), TNFα (c2), IL-6 (c3) and IL-10 (c4) in serum were examined at the indicated times after CLP by ELISA (n = 6). (d) The effects of chop-knockdown on the production of IL-1β (d1), TNFα (d2), IL-6 (d3) and IL-10 (d4) in peritoneum lavage were examined at the indicated times after CLP by ELISA (n = 6). (e) Blood was collected at 96 h after CLP and stimulated with LPS (100 ng/ml) for 1 h. The level of TNFα was examined by ELISA (n = 3). (f) The percentages of CD4⁺ and CD8⁺ T cells in the spleen were examined at 96 h after CLP by flow cytometry (n = 3). The flow histograms are presented in Supplementary Figs. S19 and S20. (g) The ratio of CD4⁺/CD8⁺ T cells in the spleen was calculated according to the corresponding percentages of CD4⁺ and CD8⁺ T cells, respectively (n = 3). The data are represented as the mean ± SD (b–g), NS > 0.05 and ***p < 0.001, Student’s t test.