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. 2017 May 25;7:2421. doi: 10.1038/s41598-017-02653-9

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© The Author(s) 2017

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 impaired capacity of activated T cell-treated fetal thymuses to support T cell development in vivo. Fetal thymuses (CD45.2⁺) were first mixed with naive or activated T cells in hanging drops for 24 hours and then cultured in FTOC for 12 days before being implanted under the kidney capsules of CD45.1⁺ mice. The development of host T cells in the implants was analysed by flow cytometry. (a) Representative CD4/CD8 staining profiles of CD45.1⁺ host-derived thymocytes at different time points after transplantation. The number indicates the percentage of cells in each quadrant. (b) Cell counts of host-derived thymocytes. (c) The percentage of DP thymocytes. (d) The absolute number of DP thymocytes. The experiments were repeated three times with at least 5 mice for each experiment. Data are presented as Mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001; and ns, not significant. In addition to the point-to-point comparisons shown in the figure, two-way Anova analysis was performed to determine the statistical significance of the differences between the two groups over the time course in terms of total thymocyte number (p < 0.0001), percentage of DP thymocytes (p < 0.0001) and absolute number of DP thymocytes (p < 0.0001).