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. 2020 Mar 24;10:5343. doi: 10.1038/s41598-020-61683-y

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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 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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Flow cytometric analysis of spleen cells at 24 hr post IPL CpG-ODN or saline treatment. (A) Macrophage analysis was performed by gating on monocytes/macrophages population in the spleen based on forward and side scatter. Bar diagram represents percent monocytes/macrophages infiltration. (B) Histogram and bar diagram displays the median fluorescence intensity (MFI) of CD40 (upper panel) and MHC II (lower panel) expression on monocytes/macrophages, respectively. Cells were gated as shown in the panel A. (C) T lymphocyte population in the spleen analysed by gating on the lymphocyte population based on forward and side scatter plot. Quantification of CD4⁺ and CD8⁺ T lymphocytes was done by using PE-labeled mouse anti-chicken CD4 and FITC-labeled mouse anti-chicken CD8 monoclonal antibodies. Bar diagram indicates the mean total CD4⁺ plus CD8⁺ T lymphocyte percentages in the spleen following IPL CpG-ODN treatment. Difference of cellular infiltration and monocyte/macrophages maturation marker expression (MFI) between the CpG-ODN administered group vs saline control group were compared using Student-t test with Mann-Whitney test, with P < 0.05 being statistically significant. Error bar = Standard Deviation (n = 4).