Network and density of lymphatic vessels in PC-3 prostate tumors from alendronate-treated and PBS-treated mice. PC-3 cells were analyzed by Northern and Western blots for expression of VEGF-C (A, B, respectively, from left to right: culture media from PC-3 cells, positive control [culture media from MCF-7 cells stably transfected with VEGF-C; n = 1 × 106 cells], negative control [normal goat IgG]). Sections from PC-3 prostate tumors were immunostained for lymphatic endothelial cells using m-LYVE-1 antibodies (C, D, E). Positive staining was observed primarily in the tumor periphery and in the peritumoral area. Tumor cells were observed also inside lymphatic vessels (E, arrows). Densities of LYVE-1-positive vessels were analyzed from the 3 best vascularized fields/tumor n (ALN) = 11, n (PBS) = 13. No statistically significant difference was detected between alendronate-treated and control tumors (F, p = 0.67, Student's t-test). (Bar = 100 μm).