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. 2018 Nov 15;8:16905. doi: 10.1038/s41598-018-34897-4

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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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The phenotype of bursal B cells rapidly changes at around hatching. Single cells produced from the bursa were stained with anti-Bu-1 antibody together with anti-IgY, -IgM or -MHC class II antibody. The percentage and absolute number of (a) IgY⁺ and (b) IgM⁺ bursal B cells before and after hatching was examined using flow cytometry. (c,d) Surface expression of MHC class II on bursal B cells was classified as a negative (−), low (+) and high (++). (c) Expression of MHC class II in bursal B cells at indicated stage. The number in each area indicates the percentage of cells. (d) The percentage and absolute number of MHC class II expression in bursal B cells during the embryonic and post-hatching stages. (e,f) B cells were distinguished by cell size as small (low forward-scatter) or large (high forward-scatter) using flow cytometry. (e) Differential display of small or large bursal B cells at indicated stage. Numbers in each area show the percentage of small or large cells. (f) The percentage and absolute number of small or large bursal B cells during the embryonic and post-hatching stages. Each data point represents the mean of four to ten chickens with SD.