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. 2017 Nov 21;7:15928. doi: 10.1038/s41598-017-15918-0

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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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Mature SP thymocytes are reduced in NCOR1 cKO^Cd4 mice. (a) Flow cytometry analysis of CD4, CD8α, TCRβ, CD24 and FOXP3 expression in WT and NCOR1 cKO^Cd4 thymocytes. The middle panel shows CD24 and TCRβ expression on CD4SP and CD8SP thymocytes. (b) Percentages and cell numbers of all thymocytes, DP, CD4SP and TCRβ^hi CD8SP thymocytes, as well as percentages of FOXP3⁺ T cells within the CD4SP subset in WT and NCOR1 cKO^Cd4 mice. (c) Bar diagrams show the relative distribution of CD24^hi and CD24^lo cells within the CD4SP and TCRβ^hi CD8SP subsets. (d) Expression of CD4 and CD8α on thymocytes isolated from OT-II,WT and OT-II,NCOR1 cKO^Cd4 littermates. (e) Flow cytometry analysis of B220 and TCRβ expression on splenocytes isolated from OT-II,WT and OT-II,NCOR1 cKO^Cd4 littermates. Histograms depict Vα2 expression on TCRβ⁺ CD4⁺ OT-II T cells. (f) Histogram depicts EGR2 expression in immature (TCRβ^hiCD24^hi) thymocytes of WT and NCOR1 cKO^Cd4 mice. WT TCRβ^−/lo CD24^hi DP thymocytes were used as a negative staining control. (g) Histograms depict CD5 expression on DP (upper panel), CD4SP (middle panel) and TCRβ^hi CD8SP (lower panel) thymocytes in WT and NCOR1 cKO^Cd4 mice. (a,d,e) Numbers indicate the percentage of cells in the respective quadrants. (b) Thick horizontal bars indicate the mean. (b,c) *P < 0.05, **P < 0.01 and ***P < 0.001 (unpaired two-tailed Student’s t-test). Data are representative (a,d,e,f,g) or show the summary (b,c) of 7–10 mice (a,c), 25–26 mice (b; except for CD4SP FOXP3⁺ cells, where n = 7 mice) and 5–6 mice (d,e,f,g) that were analyzed in 3–4 (a,c), 8 (b; except for CD4SP FOXP3⁺ cells: 3), 3 (d,e,g) and 2 (f) independent experiments.