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. 2018 Oct 2;8:14676. doi: 10.1038/s41598-018-33101-x

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© The Author(s) 2018

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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Effects of LPS or GYY4137 on Hoxb8 neutrophil vitality and migration through an endothelial monolayer in vitro. Differentiated neutrophils were incubated for 4 h in the absence (−) or presence (+) of GYY4137 as indicated, and the relative amount of vital cells was detected by propidium iodide staining and FACS analysis (a). Differentiated neutrophils were placed onto HUVEC endothelial monolayers in the upper compartment. Cell culture medium was either left untreated in all compartments (control) or was supplemented with 1 mM GYY4137 in the upper compartment (control + GYY). Cell culture medium in the lower compartment was supplemented with MIP-2, either in the absence (MIP-2) or presence of 1 mM GYY4137 in the upper compartment (MIP-2 + GYY, b). After incubation for 2 h, cells from the lower and upper compartment were stained separately with propidium iodide and analysed by FACS. The relative amount of migrated neutrophils was calculated as the ratio between vital cells in the lower compartment and the total number of vital cells (c). Graphs represent means ± SEM, n = 3/group. ANOVA (Tukey’s post hoc test), *GYY vs. control (c: P = 0.0030); *MIP-2 vs. control (c: P = 0.0039); ^#MIP-2 vs. control + GYY (c: P < 0.0001); ^§MIP-2 + GYY vs. MIP-2 (c: P = 0.0002).