4.3. Generation of A375 Melanoma Xenografts

All animal experiments were carried out according to the guidelines of the German Regulations for Animal Welfare. The protocols were approved by the local Ethical Committee for Animal Experiments (AZ DD24.1-5131/449/49). Generation of tumor xenografts was performed as described elsewhere [43]. In brief, Rj:NMRI-Foxn1 nu/nu mice were s.c. injected with 5 × 10⁶ A375-pIRES and A375-EphB4 cells, each in 100 μL 0.9% v/v NaCl, into the left and right hind leg, respectively. Tumor size was monitored thrice a week by caliper measurements and tumor volume was calculated using the formula V = π/6 × (tumor length × tumor width²). Tumor-bearing mice were included into the experiments about 14 days post tumor cell injection, when tumors reached a volume of at most 1000 mm³.