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. 2017 Jun 21;7:3974. doi: 10.1038/s41598-017-04322-3

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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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PSTreg show specific immunosuppressive activity. Three different assays were used to test PSTreg suppressive function towards PSTeff. PS/NTreg and PS/NTeff were seeded in 96 U-well plates with CD3/CD28 beads at different ratios, NTeff and PSTeff were set up as controls. Anti-CD154 and anti-CD25 were used to determine the Teff activation suppression, and CellTrace™ CFSE Cell Proliferation Kit was used to determine the inhibition of Teff proliferation. (a) Suppression of CD154 after 7 h at different ratios. (c) Suppression of CD25 after 96 h. (e) Inhibition of Teff proliferation at different ratios. (n = 3). (b) Suppression of CD154 at ratio 1:1, (d) shows suppression of CD25 at ratio 1:1, Data represents 15 independent experiments from 8 donors and 10 independent experiments from 6 donors, respectively. (f) Inhibition of Teff proliferation at ratio 1:1, (e) Data represents 8 independent experiments from 5 donors. ***p < 0.001, **p < 0.01, *p < 0.05. NS, not significant (p > 0.05). ANOVA with Bonferroni’s Multiple Comparison Test was used to determine the statistical significance. Error bars: s.e.m.