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. 2017 Jun 23;117(8):1727–1737. doi: 10.1007/s00421-017-3667-0

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© The Author(s) 2017

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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.

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Fig. 1 — Gating strategy to define Treg subsets. A Gating was performed to exclude debris (a), doublets (b), red blood cells (c) and dead cells (d) using time during acquisition, forward and side scatter properties and viability dye zombie NIR. Final gating is on the lymphocyte population (e). B Gating strategy to define Treg subsets within the CD3⁺CD4⁺ population (f): CD25⁺⁺Foxp3⁺ (g) and CD127⁻ total Tregs (h); CD45RA⁺ Tregs (i) and HLA-DR⁺ Tregs (j)