Male nude mice

Male nude mice (5–6 weeks old) were purchased from Taconic and maintained and treated under specific pathogen-free conditions. All procedures were approved by the Institutional Animal Care and Use Committee at Beth Israel Deaconess Medical Center and conform to the federal guidelines for the care and maintenance of laboratory animals. The mice were injected subcutaneously with 9 × 10⁶ LNCaP cells in media with 50% Matrigel. Tumor formation was examined every 2 to 3 days for the duration of the experiment. Mice with palpable tumors were randomly divided into control and treatment groups and treated with water supplemented with 1 mg ml⁻¹ dox and 2% sucrose, or standard water supply for 12 days. Bottles of water/dox were changed twice a week. Tumor volume was measured on the first day of dox treatment, as well as day 12 post-dox treatment. Tumor volume = (4/3)(π)(R1 × R2 × R3); R = radius. Tumors were harvested at the experimental endpoint.

Xenografts were established in the flanks of male nude mice (Taconic) by subcutaneous inoculation of ~3 million of indicated cells mixed with 50% Matrigel (BD Bioscience). When the tumors reached the indicated volumes, treatments were initiated. Growth was then either monitored by caliper measurements or tumor samples were collected for protein analysis or IHC staining. All animal experiments were approved by Beth Israel Deaconess Medical Center’s Institutional Animal Care and Use Committee and were performed in accordance with institutional and national guidelines.