Figure 7: TAMs mediate TNF-α-lymphangiogenesis and metastasis.

(a) FACS analysis of the F4/80⁺ pan population and F4/80⁺CD206⁺ M2 subpopulation of TAMs in vector and TNF-α tumours (n=6 samples per group). (b) Immunohistochemical analysis of F4/80⁺ (red) and CD206⁺ TAMs (green) in TNF-α tumour-bearing wt and Tnfr1^−/− mice. Vector tumours implanted in wt mice served as a control. Scale bar, 100 μm. Quantification data were obtained from eight randomized fields per group. (c) Confocal images of TAMs and lymphangiogenesis in primary tumours. TAMs (red) and tumour lymphatic vessels (green) in clodronate liposome-treated TNF-α tumour-bearing wt mice. TAMs and lymphatics in TNF-α tumour-bearing Tnfr1^−/− mice were also analysed. Vector tumours served as a control. Arrowheads point to tumour lymphatic vessels. Scale bar, 100 μm. Quantification of tumour TAMs and LYVE-1⁺ lymphatic vessels in various treated and non-treated groups. Quantification data were obtained from eight randomized fields per group. (d) Morphology (scale bar, 1 cm) and hematoxylin and eosin stain (H&E) histological staining (scale bar, 100 μm) of LNs from various treated and non-treated tumour-bearing mice. Dashed lines mark the border between metastatic tumour and LN tissues. T, tumour. (e) Quantification of LN volume and weight of various groups (n=6 mice per group). *P<0.05; **P<0.01; ***P<0.001. All error bars represent s.e.m. All P values were analysed according to Student’s t-test.