Glioblastoma Cell Culture and Treatments

T98G, A172, LN229, MCF-10A and HMC3 human microglial cells were from ATCC; U251-MG and SF188 cells were a gift from Dr Chris Jones (ICR, London, UK). GBM cell lines were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (cat#11995065, Gibco), HMC3 cells in DMEM/F12 (cat#10565018, Gibco), and MCF-10A cells in mammary epithelial cell basal medium (MEBM) (cat#CC-3151, Lonza). Human astrocytes (cat#3P10251; Innoprot) were cultured in AM (cat#1801, ScienCell) with supplements (cat#1852, ScienCell). Mouse GSCs were isolated and cultured as described previously [47 (link)]. For radiation, cells were irradiated with a linear electron accelerator (Varian Medical Systems) using a 6MV photon field at varying doses ranging from 6−12 Grays. Dexamethasone (cat#D4920), temozolomide (cat#T2577), human PDGF-BB (cat#P3201) and LPS (cat# L5293) were from Sigma/Merck (Madrid, Spain). IFNγ (cat#300-02) was from Peprotech (London, UK) and reversine (cat#ab120921) from abcam (Cambridge, UK). FDA approved drug library (cat# L1300), Ponatinib (cat#S1490), Sunitinib (cat#S7781), Dasatinib (cat# S1021), Venetoclax/ABT-199 (cat# S8048), S63845 (cat# S8383), Vincristine (cat#S1241) and Taxol (cat#S1150) were from Selleckchem (Newmarket, UK). Navitoclax/ABT-263 (cat#M1637) was from AbMole. TKIs were used at concentrations in the range of their IC50s (see Figure S5A).