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. 2023 Feb 4;12(4):516. doi: 10.3390/cells12040516

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© 2023 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Gating strategy for the assessment of CD8+ and CD4+ CD45RA+ CCR7- effector memory T cells (T_EMRA) and their subsets. T cells were gated as CD3+ leukocytes and divided into CD8+ and CD4+ T cells. Both CD8+ and CD4+ T cells were divided into naive T cells (T_N), central memory T cells (T_CM), effector memory T cells (T_EM), and effector memory T cells re-expressing CD45RA (T_EMRA) based on the expression of CD45RA and CCR7. T_EMRA cells were defined as CD45RA+ CCR7-. Both CD8+ and CD4+ T_EMRA cells were divided into further subsets based on the expression of CD27 and CD28 and on the expression of CD28 and CD57. Gates for CCR7, CD45RA, CD27, and CD57 were set using fluorescence minus one (FMO) controls.