Fig. 3

Analysis of the tumoral vasculature, proliferation and necrosis of liver metastases of WT31 and WT31_P5IV. A Images of immunofluorescence staining for Emcn (green), Lyve-1 (red) and TRP-2 (blue) of liver metastases of WT31 and WT31_P5IV. Representative images are shown. Scale bars = 200 µm, n = 4. White dotted lines present the border of hepatic metastasis of WT31 and WT31_P5IV to the adjacent liver tissue. B Images of immunofluorescence staining for CD31 (green) and CD32b (red) of liver metastasis of WT31 and WT31_P5IV. Representative images are presented. Scale bars = 200 µm, n = 5. White dotted lines present the border of hepatic metastasis of WT31 and WT31_P5IV to the adjacent liver tissue. C Intratumoral endothelial cells were analyzed for Emcn or Lyve-1 marker expression. The corresponding marker expression was set in relation to the total intratumoral vessel area (100%). The proportion of Lyve⁺, Lyve⁺  + Emcn⁺ as well as Emcn⁺ area is displayed (Emcn: P > 0.9999; Lyve-1 and Emcn: P = 0.7000; Lyve-1: P > 0.9999; Mann–Whitney-tests). Besides, the expression of CD31 and CD32b of intratumoral vessels was analyzed and set in relation to the total intratumoral vessel area (100%). The proportion of CD31⁺, CD31⁺  + CD32b⁺ as well as CD32b⁺ area is displayed. (CD31: P = 0.6325; CD31 and CD32b: P = 0.7118; CD32b: P = 0.6492; t-tests). Last, the vascular density of the hepatic melanoma metastases was determined. The CD31.⁺ area in relation to the total tumor area is shown (P = 0.3776, t-test) D Images of immunofluorescence staining for CD31 and Ki67 of liver metastases of WT31 and WT31_P5IV. Representative images are presented. Scale bars = 200 µm, n = 6. White dotted lines present the border of the hepatic metastasis of WT31 and WT31_P5IV to the adjacent liver tissue (P = 0.6282, Mann–Whitney test). E Images of immunofluorescence staining for CD31 and cCasp3 of liver metastases of WT31 and WT31_P5IV. Representative images are shown. Scale bars = 200 µm, n = 4. White dotted lines present the border of the hepatic metastasis of WT31 and WT31_P5IV to the adjacent liver tissue (P = 0.1163, t-test)