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. 2019 Sep 25;7(18):e14234. doi: 10.14814/phy2.14234

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© 2019 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of The Physiological Society and the American Physiological Society.

This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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Representative gating procedures for analyzed samples. (A) Illustrates usage and placement of the live cell gate in the forward scatter versus side scatter plot. Sample populations were all >97% CD3⁺CD4⁺ following isolation. All flow data were gated as in (A) before further analysis. An unstained sample (not shown) was used as a guide for placement of the CD4⁺ gate in the fully stained sample (B). Expression of surface markers of activation in non‐stimulated and stimulated cell populations were quantified using median fluorescent intensity (MFI) of (C) CD69 and (D) CD25. Overlays of the non‐stimulated (light gray) and stimulated samples (dark gray) were used to correct for the effect of costimulation through CD28 prior to analyzing exercise‐induced alterations.