(A) Normalized gene expression of thrombospondin-1 (TSP-1) in the R-18, A-07, and D-12 melanoma model. Columns, mean of three independent experiments; bars, SEM. (B) TSP-1 concentration in the blood plasma of untreated mice bearing R-18, A-07, or D-12 tumors. Columns, mean of 6 mice; bars, SEM; N.D., not detectable. (C) Immunohistochemical preparations of untreated R-18, A-07, and D-12 tumors stained with anti-TSP-1 antibody. Scale bars, 25 μm. (D) Normalized gene expression of vascular endothelial growth factor A (VEGF-A), interleukin 8 (IL-8), basic fibroblast growth factor (bFGF), and platelet-derived endothelial cell growth factor (PD-ECGF) in the R-18, A-07, and D-12 melanoma model. Columns, mean of three independent experiments; bars, SEM. Gene expressions were measured with quantitative PCR, was normalized to the mean expression of two housekeeping genes (GAPDH and ACTB), and multiplied with 100 (A and D).