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. 2020 Apr 30;18:183. doi: 10.1186/s12967-020-02348-2

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© The Author(s) 2020

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 2 — Macrophages within the metastatic niche are shifted to pro-tumor state in the acute post-operative period. a–c Mice were treated as in Fig. 1g, then euthanized 48 h after surgical interventions (9 days after tumor implantation) and lungs processed into single cell suspensions. Flow cytometric analysis of lungs from tumor-bearing mice (n = 10), mice that underwent surgical excision of their tumor-bearing limb (amputation, n = 10), or mice that underwent surgical excision of their contralateral, non-tumor bearing-limb (n = 10) were compared via flow cytometric analysis. After gating for viability, cells were gated for size, singlets, and CD45, F4/80 double positivity, to define the general macrophage population. CD45⁺/F4/80⁺ cells were analyzed for the percentages of surface markers CD206 and MHCII. a Percentage of MHCII⁺/CD206⁻ macrophages (anti-tumor macrophage population). b Percentage of/MHCII⁻/CD206⁺ macrophages (pro-tumor macrophage population). c Pro-tumor macrophages: anti-tumor macrophage ratio. Bars represent mean ± standard deviation compared by one-way ANOVA with post hoc Dunnett’s multiple comparisons test **p < 0.01