Fig. 3

Gefitinib treatment prevents surgery-accelerated metastasis and reverses post-surgical pro-tumor macrophages polarization. a BALB/c mice were implanted with 3 × 10⁵ cells in the left tibia and 1 week after tumor implantation mice underwent surgical excision of primary tumor bearing limb or amputation of the contralateral limb as in Fig. 1g. Mice assigned to gefitinib treatment were started on gefitinib-impregnated chow (100 mg/kg/day) 2 days prior to surgical excision (5 days after tumor implantation). Gefitinib treatment was continued for 1 week following surgery, and mice were sacrificed 3 weeks after surgery (4 weeks after tumor implantation). a Number of gross nodules on surface of lungs (n = 7–10 mice per group). b Mice were treated as in Fig. 1. Lungs were made into single-cell suspensions and analyzed via flow cytometric analysis with gating strategy as in Fig. 2. Combined data from three independent experiments (n = 4–6 mice per group per experiment). Pro-tumor macrophage: anti-tumor macrophage ratio. Scatter plots show individual mice, all graphs displayed with means ± standard deviations. Data analyzed using one-way ANOVA with post hoc Tukey’s multiple comparison test,*p < 0.05, **p < 0.01