Nmri nu

1.2 × 10⁶ WM266-4 cells were subcutaneously injected into the flank of 6 weeks female athymic mice (NMRI-NU, Janvier) and allowed to grow 10 days. Subsequently, animals received by oral gavage 50 mg/kg/bid of PLX4032 and/or 20 mg/kg/day G594 or vehicle. PLX4032, formulated as MBP, and G594 were provided by Genentech and prepared according to their advices. Tumor sizes were evaluated every 3 days by caliper measurement of two perpendicular diameters (l < L) and the tumor volumes were calculated with the following equation tumor volume = l × l × L/2. Animals were euthanized after 21 days of treatment, tumor were harvested and weighted. All animal procedures were performed in accordance with the animal care guidelines of the European Union and French laws and were approved by the local Ethical Commission of the Institut Claudius Regaud.

For the grafting protocol immunodeficient Rj: NMRI-Foxn1nu (NMRI nu) female mice (6 weeks old) were used (Elevage Janvier, France). Mice were aseptically cleansed and grafted. They were housed for the duration of the experiment at the CIEMAT Laboratory Animal Facility (European Registration number ES280790000183; approved date: 17 April 2012) in pathogen-free conditions. All experimental procedures were carried out according to the European and Spanish laws. All procedures were approved by the Institutional Animal Care and Use Committee according to all legal regulations, and biosafety and bioethics guidelines. During the experiments, a minimum number of n = 3 animals were studied for each condition described in the results section.