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. 2020 Jan 17;10:3109. doi: 10.3389/fimmu.2019.03109

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Copyright © 2020 Zhu, He, Liu, Qu, Xie, Yang, Lei, Zhou, Shi, Zhang, Cheng, Sun, Zheng, Shen, Li, Jiang and Zhang.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Microbiota antigens stimulate the expansion of eTregs. Purified tdTomato⁻CD4⁺ T cells were labeled with CTB dye and transferred to either sublethal irradiated WT mice or Rag2 KO mice. Four days post cell transfer, tdTomato expression in the splenocytes of the recipients was analyzed. (A) FACS plots for the percentage of tdTomato⁺ cells in subpopulations of donor cells. (B,C) FACS plots and histogram of Foxp3 expression in tdTomato⁺CD25⁺ cells from RAG2^−/− group. (D) The percentage of tdTomato⁺ Tregs in indicated groups. (E) Statistics for tdTomato⁺ cells in Tregs from two groups (n = 4). Each plot represents three independent animals. The experimental procedure for (D,E) was described in Methods. **P < 0.01.