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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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Large B cells are the main source of proliferation and differentiation in the embryonic stage. To explore whether the proportion of small and large B cells affects the status of B cells during the development, we chose two stages where the ratio of small and large B cells are dramatically reversed. Cells in the large B cell dominant stage (E18) and a small B cell dominant stage (E20) were sorted using Bu-1 positive selection, and then total RNA was extracted. (a–h) The mRNA expression of B cell proliferation- and colonization-associated genes in small and large B cells was evaluated using qRT-PCR as follows: (a) BAFF, (b) BAFF receptor, (c) IL-7, (d) IL-7 receptor, (e) CD40, (f) CD40 ligand, (g) CXCL12, and (h) CXCR4. (i–s) The mRNA levels of B cell differentiation-associated genes in small and large B cells were examined using qRT-PCR as follows: (i) RAG1, (j) RAG2, (k) FOXO1, (l) FOXO3, (m) PAX5, (n) BLNK, (o) Ikaros, (p) Helios, (q) Aiolos, (r) EBF, and (s) E2A. *P < 0.05, **P < 0.01, ***P < 0.001.