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