Nu nu mice

Xenograft experiments were approved by the University of Colorado Institutional Animal Care and Use Committee (IACUC protocol 83614(01)1E). All animal experiments were conducted in accordance with the NIH Guidelines of Care and Use of Laboratory Animals. A total of 10⁶ SUM159PT-TGL or 500,000 HCC1806-TGL cells were mixed with Matrigel (BD Biosciences) and bilaterally injected into the mammary fat pads of female, athymic nu/nu mice (Taconic). Tumor burden was assessed by luciferase activity and caliper measurements [tumor volume was calculated as volume = (length × width²)/2]. Once tumors were established, mice were randomized into groups based on the total tumor burden as measured by in vivo imaging. Mice were administered enzalutamide in their chow (~a 50 mg/kg daily dose). Enzalutamide was mixed with ground mouse chow (Research Diets Inc.) at 0.43 mg/g chow. The feed was irradiated and stored at 4°C before use. Mice in the control group received the same ground mouse chow but without enzalutamide. All mice were given free access to enzalutamide-formulated chow or control chow during the study period. Mice were euthanized by carbon dioxide asphyxiation followed by cervical dislocation, and the tumors and mammary glands were harvested.

Five million DIRAS3-inducible cells and the parental OVCAR8 cell lines were injected intraperitoneally into six-week old female athymic nu/nu mice were purchased from Taconic Biosciences. Each group contained 8-10 mice. 2 mg/mL DOX in 5% sucrose or sucrose alone water starting the day of injection, and supplemented every two days throughout the duration of the study. The tumor mass (g) was determined 4 weeks post injection after euthanizing mice with CO₂ and excising and pooling each intraperitoneal tumor nodule. All procedures were carried out according to an animal protocol approved by the IACUC of The University of Texas MD Anderson Cancer Center.

The animal work was in accordance with a UCSF Institutional Animal Care and Use Committee protocol. Six to seven-week-old nu/nu mice were purchased from Taconic Farms. Nude mouse xenografts were generated by subcutaneous injection of each cell line (1 × 10⁶ cells/ml; 100 μl per site/mouse). Animals for imaging and biodistribution studies had tumor volumes between 100 – 350 mm³. The intracardiac dissemination model was generated using the previously described method (35 ).

All animal experiments were approved by the Institutional Animal Care and Use Committee at Washington University in St. Louis School of Medicine. To generate tumor xenografts, 8-week-old nu/nu mice (Taconic) were injected subcutaneously in the flank with 3 × 10⁶ HT1080 cells in 100 µL αMEM. Tumors were allowed to grow for 11 days before treatments were started. Intratumoral injection was performed on days 11, 13, and 15 with 20 µL of solution containing vehicle (DMSO) or Zaprinast (600 µM) (Sigma-Aldrich). Tumor volume was calculated as l × w × h of tumor dimensions obtained by caliper measurements. On day 15, tumors were extracted and snap frozen for follow up GC-MS analysis.