Ln 319

Human GIC (S-24, T-269, T-325, ZH-161, and ZH-305) were established from freshly resected tumors [62 (link)]. The human long-term glioma cell lines LN-18, LN-428, D247MG, LN-319, A172, LN-308, and LN-229 [63 (link)] were kindly provided by N. de Tribolet (Lausanne, Switzerland) and T98G cells were obtained from the American Type Culture Collection (ATCC) (Rockville, MD). GIC were cultured as neurospheres in Neurobasal medium (NB) supplemented with 2 mM l-glutamine, 20 μg/ml B-27 supplement (Gibco, Waltham, MA), 20 ng/ml fibroblast growth factor (FGF) 2, and 20 ng/ml epidermal growth factor (EGF) (Peprotech, Rocky Hill, PA). Long-term glioma cell lines were grown as adherent monolayers in Dulbecco´s modified Eagle´s medium (DMEM) supplemented with 10% fetal calf serum (FCS) and 2 mM l-glutamine (Gibco). Cells were regularly tested for mycoplasma contamination by means of MycoAlert^TM PLUS Mycoplasma Detection (Lonza, Basel, Switzerland).

The human glioma cell lines LN-18, LN-229, LN-319, T98G, U251, U138MG and LN-428 were purchased from the American Type Culture Collection (Manassas, VA, USA). The human glioma cell line LN-308 was provided by N. de Tribolet. Regular checks for authentity and freedom from infection, e.g. mycoplasms, were done according to the institutional guidelines at the German Cancer Research Center. Details concerning cloning, quantitative reverse transcription PCR analyses (including Table S3), sequencing (including Table S4), shRNA constructs for VEGFR-2, siRNA constructs for Raptor or Rictor and overexpression constructs for VEGFR-2 or PTEN including the respective controls are given in the Supplementary Information.