Nu nu female mice

All animal procedures were approved by the Fred Hutchinson Cancer Research Center Institutional Animal Care and Use Committee. Nu/Nu female mice 4 weeks old were purchased from Envigo (East Millstone, NJ, USA). SB01-VR xenografts were established by subcutaneous injection of 5×10ˆ6 cells into the right flank. Five days after injection, mice were randomized into groups of 10 to obtain groups with similar starting average tumor size. Groups of 10 mice allowed us to observe tumor size differences of 15% or more that are 1.1 standard deviation units with 80% power in a 2-sided test with 0.05 level of significance. Bafetininb, trametinib, combination bafetinib and trametinib, or vehicle (0.5% methylcellulose, 5% DMSO, 0.2% Tween 80) was administered by oral gavage for 21 consecutive days. Bafetinib was dosed at 200mg/kg/d (100mg/kg twice daily); trametinib was dosed at 0.1mg/kg once a day^{34 (link), 35 (link)}. Mice treated with drug combinations were dosed with 100mg/kg bafetinib plus 0.1mg/kg trametinib in the morning and only 100mg/kg bafetinib at night. Control mice were also dosed twice daily. Tumor sizes were assessed three times a week by caliper measurement and volumes calculated from the formula, tumor size (cm3)=(L×Wˆ2×3.14159/6) using Study Advantage Tumor Tracker software (Jenkintown, PA, USA).