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. 2016 Jul 11;6:29601. doi: 10.1038/srep29601

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Copyright © 2016, Macmillan Publishers Limited

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(A) Flow cytometric analysis of blood neutrophils and monocytes/macrophages. Monocytes/macrophages were identified as CD11b^highLineage^lowLy6C^high/low, and neutrophils were identified as CD11b^highLineage^high. Lineage refers to the combination of CD90, B220, CD49b, NK1.1 and Ly6G monoclonal antibodies. (B) Representative histograms from individual mice within the CD11b^high Myeloid cells gate depict neutrophils and monocytes/macrophages in healthy hearts and within the heart at specified times after IR. Column graphs illustrate the total number of neutrophils per heart (N = 4 mice per group). (C) Representative histograms from individual mice within the monocytes/macrophages cell gate depict Ly6C^high and Ly6C^low monocytes in healthy hearts and within the heart at specified times after IR. Column graphs illustrate the total number of monocytes/macrophages per heart (N = 4 mice per group). (D) The percentage of Ly6C^high monocytes/macrophages and the total number of Ly6C^high monocytes/macrophages per heart. (E) The percentage of Ly6C^low monocytes/macrophages and the total number of Ly6C^low monocytes/macrophages per heart. (F) Flow cytometry of leukocytes in the healthy hearts and hearts 48 h after IR injury. Red gate shows the population of macrophages (CD11b^highLineage^lowF4/80^highLy6C^low cells). The percentage and the number of macrophages, Ly6C^high monocytes and Ly6C^low monocytes per CD11b^highLineage^low cells in IR heart. Data are expressed as the mean ± SEM (N = 4 mice). All results are average of three replicates. IR: ischemia reperfusion, FCM: flow cytometry, Mφ: macrophage, Mono: monocyte.