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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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In vivo derived DCs expand Tregs through OX40L. (A) Representative dot plots of OX40L⁺CD11c⁺ expression within various tissues of untreated and GM-CSF treated mice (n = 3). (B) Bar graphs show percent OX40L⁺CD11c⁺ DCs in indicated tissues. Values are expressed as means ± SEM (n = 3; *p < 0.05, **p < 0.01). (C) Representative histograms of indicated markers on OX40L⁺CD11c⁺ (black) compared to OX40L⁺CD11c⁻ (grey) isolated cells. Dotted lines represent stained controls. (D) Bar graphs showing the expression of indicated markers on OX40L⁺CD11c⁺ DCs and OX40L⁻CD11c⁺ DCs. Values are expressed as means ± SEM (n = 3; **p < 0.01, ***p < 0.001). (E) CD11c⁺ DCs were isolated from the spleens of GM-CSF treated or WT mice and co-cultured with CD4 T-cells with or without OX40L blockade and/or supplemented with OX40 agonist. Representative dot plots are shown. (F) Summary bar graphs showing percent proliferating Foxp3⁺ Tregs in indicated co-cultures. Values are expressed as means ± SEM (n = 3; **p < 0.01, ***p < 0.001).