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. 2021 Jul 22;11:14986. doi: 10.1038/s41598-021-94016-8

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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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Neutrophil depletion increases the proportion of CD11b⁺Ly6C^hi cells in the liver after IRE and shifts the macrophage population toward a more pro-inflammatory state. (a) Neutrophil depletion protocol and timeline in which rat IgG2a (isotype control) or anti-Ly6G were given as 1° antibodies and anti-rat IgG given as the cross-linking 2° antibody. The timing of IRE treatment and analysis of liver myeloid cells are indicated by arrows. (b) Percentage of neutrophils in the blood at baseline (prior to any IRE or antibody treatments), and on days 0, 1, 3, and 7 after IRE treatment with the neutrophil depletion regimen shown in (a). (c) Percentage of neutrophils in IRE-treated liver parenchyma 3 days after treatment in mice given isotype control or anti-Ly6G. (d) Total CD11b⁺ cells or CD11b⁺Ly6C^hi cells as a percentage of CD45⁺ leukocytes in IRE-treated liver parenchyma 7 days after treatment in mice with or without neutrophil depletion. (e) Ly6C^hi to Ly6C^lo ratio in IRE-treated liver 7 days after treatment in mice given isotype control versus anti-Ly6G. For all graphs, unpaired two-tailed t tests show *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. Data points represent individual animals in each condition and/or timepoint (n = 3–8). Graph bars and error bars show mean ± SEM.