Irradiator gamma cell exactor 40

Investigation has been conducted in accordance with the ethical standards according to national and international guidelines and has been approved by the national animalcare committee. 5×10⁶ cells were inoculated subcutaneously into 4 to 5 weeks-old athymic nude mice (Charles River, France). Tumors sizes were measured twice a week with a digital calliper. When the average tumors volume was about 150-200 mm³, Animals were randomized into 4 groups (control-shRNA, PG-shRNA, control-shRNA plus radiation, PG-shRNA plus radiation; 6 animals/group). Tumours were locoregionally irradiated with fractionated radiotherapy, 2.5 Gy/fraction every 3 days with a total dose of 10 Gy using gamma rays (Irradiator Gamma-cell Exactor 40, Nordion, Ottawa, ON, Canada).

GC1 and GC2 neurospheres were dissociated and transfected with a control siRNA and siRNA targeting specifically FGFR1, FOXN3 or FOXM1. A day later, cells were seeded at a concentration of 100 cells/well in 96 well plates and irradiated at different doses (2, 4, and 6 Gy), using an Irradiator Gamma-cell Exactor 40 (Nordion, Ottawa, ON, Canada). After irradiation, survival cells constituted neurospheres. Neurospheres were counted when they are composed of at least 20 cells.