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. 2018 Oct 8;8:14940. doi: 10.1038/s41598-018-33307-z

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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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OX40L⁺CD11c⁺ G-BMDCs are responsible for the expansion of functional Tregs. Splenic C57BL/6 CD4⁺ T-cells were stained with CellTrace and separately co-cultured with anti-CD3/CD28 microbeads, bulk G-BMDCs (pre-sort) population, sorted OX40L⁻CD11c⁺ G-BMDCS, and sorted OX40L⁺CD11c⁺ G-BMDCS. (A) Representative dot plots of percent proliferating Tregs using CD4⁺FOXP3⁺ as a marker for Tregs. All flow cytometry plots were gated on CD4⁺ cells. (B,C) Bar graphs showing percent proliferation of Tregs and Teff in CD4⁺ T-cell co-cultures with indicated cell populations, (n = 3). The data show the means ± standard error of the mean (SEM). A p-value < 0.05 was considered significant; **p < 0.01, ***p < 0.001. (D) Representative dot plots of Treg suppressive markers on proliferating Tregs. (E) Bar graphs showing percent expression of Treg suppressive markers on proliferating Foxp3⁺ Tregs. Values are expressed as means ± SEM (n = 3; ***p < 0.001).