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. 2014 Apr 11;9(4):e94492. doi: 10.1371/journal.pone.0094492

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© 2014 Awad et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Autologous NK cells and eosinophils were cultured at the ratio NK cells: eosinophils (NK: Eos ratio) 0∶1 (i.e. eosinophils alone), 1∶1, 5∶1 and 10∶1 in the presence of interleukin-5 for 3 hours and 12 hours. (A) Gating strategy: live eosinophils were identified firstly on the basis of their size difference (FSC) and granularity (SSC) and secondly as Annexin V⁻ 7AAD⁻ cells. CD63 expression (black histogram) is shown on a representative experiment for the ratio NK cells: eosinophils (NK: Eos ratio) 0∶1 (i.e. eosinophils alone) and 10∶1. Isotypic control antibody is represented in dashed grey. Eosinophils were stained for CD63 (B), CD62L (C) and CD69 (D) expression. Results are shown for each donor and mean is indicated with the black line. One-way Anova (Friedman) tests were performed, followed by Dunn’s post tests. *p<0.05; ***p<0.001.