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. 2018 Jun 11;6(11):e13721. doi: 10.14814/phy2.13721

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© 2018 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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Gating strategy used for CD45⁺ subpopulations. Pregating on LIVE cells, CD45⁺ leukocytes were analyzed for myeloid populations (right fork, Gibbings et al. 2017) as well as neutrophils (left fork; Ly6G⁺ versus side scatter [SSC‐A], Yu et al. 2016), in mouse left lungs D1 after the onset of ischemia. Macrophages (CD64⁺ MerTK ⁺ CD11c⁺), were either alveolar macrophages (ALV; CD11b⁻) or interstitial macrophages (IM; CD11b⁺). Interstitial macrophages (IM) were further evaluated for MHCII ^{hi, intermediate, lo}. CD64⁻MerTK ⁻ cells were further defined based on F4/80 staining and side scatter (SSC‐A) with the second analysis of neutrophils (N) as F4/80^lo and eosinophils (Eos) as F4/80^intermediate and monocytes F4/80⁺. Monocyte subpopulations were either Ly6C^hi or Ly6C^lo. These plots include 65,769 live events.