Male fvb nj mice

Male FVB/N-J mice were originally purchased from Jackson Laboratory and further breeding was carried out in-house as approved by the institutional animal care and use committee. Just before the tissue enzyme activity measurements, animals of two age groups (4 weeks & 40 weeks) were sacrificed and the liver tissues were surgically excised. Immediately after the surgery, the liver tissues were homogenized in 2ml microcentrifuge tube containing hypotonic buffer using a tissue lyser (Qiagen, 5 minutes, 30Hz or 1800 oscillations/minute). Protein lysates were then used immediately for measuring catalase activity as described. In another set of experiments, 50mg fresh liver biopsy was used to measure catalase activity in the presence and in the absence of catalase-specific pharmacological inhibitor 3AT (6mM 3-Amino, 1,2,4, triazole, Sigma Aldrich, USA).

For all the animal experiments in the present study, the study protocols were approved by the Institutional Animal Care and Use Committee (IACUC) of UT MD Anderson Cancer Center. Male BALB/c nude mice, male NOD SCID gamma mice, male C57BL/6J mice, and male FVB/NJ mice (aged 4–6 weeks) were obtained from The Jackson Laboratory. For primary tumor growth assays, cells were resuspended in 100 μL PBS with Matrigel in 1:1 ratio and subcutaneously injected into both rear flanks. The volume of the s.c. xenograft was calculated as V = L × W2/2, where L and W stand for tumor length and width, respectively. For experimental metastasis assays, cells were resuspended in 100 μL PBS and injected into the left ventricle with a 26G tuberculin syringe. For drug treatment, drug solutions were delivered either intraperitonially or by oral gavage using a 20G reuseable feeding needle (Roboz Surgical Co.). Metastatic burden was detected through noninvasive bioluminescence imaging of experimental animals using an IVIS Spectrum and Biospec 7T MRI instruments at the Small Animal Imaging Facility (SAIF). To investigate the effect of drug treatment, compounds were delivered daily through p.o. Bioluminescence signals were measured using the ROI tool in Living Image software (Xenogen).