Athymic nude mice

The immunocompetent BALB/C mice and athymic nude mice were purchased from Central Lab. Animal Inc., and were raised in a specific-pathogen-free (SPF) environment with standard conditions, controlled at a temperature of 20–25°C with 40–50% humidity with a 12-h light/dark cycle. There was no specific limitation for food or water. Mice with 6–8-week-old female with body weight ranging 18–20 g were used. For the depletion of CD4 and/or CD8 T cells in immunocompetent BALB/C mice, 0.5 mg of the anti-mouse CD4 clone GK1.5 or anti-mouse CD8 clone 53–6.72 (BioXCell) mixed with 0.5 ml of phosphate-buffered saline (PBS) was injected intraperitoneally for three consecutive days. Thereafter, the depletion status was maintained with a twice-weekly peritoneal injection of the corresponding antibodies. Five nude mice were used as comparison group, and 47 BALB/C mice were used. (control; 12, CD8 depletion; 13, CD4 depletion; 13, CD4+8 depletion; 9)

Twenty female athymic nude mice of 4-week-old were purchased from Central Lab Animal Inc. (Seoul, Korea). SNU478 xenograft model mice were established by subcutaneous inoculation of 2×10⁷ cells in 100 μL of PBS. When the tumor volume reached 200 mm³, the mice were randomly divided into four groups of five mice. AZD6738 was administered orally once a day at 25 mg/kg for 3 weeks and the control group was treated with vehicle (2-hydroxypropyl-β-cyclodextrin solution) by oral gavage. Cisplatin was injected intraperitoneally twice in the first week and then once weekly for next 2 weeks. Body weights and tumor sizes were measured every other day. The tumor volume was calculated using the formula: tumor volume=[(width)²×height]/2. None of the mice was killed during experiment by mistake and all mice were sacrificed finally.