Gibco b 27 serum free supplement

All GBM tumor samples and subsequent cell line derivations were performed with written consents from patients through Quiñones laboratory, observing Institutional Review Board guidelines as previously described. GBM stem cells with tumorsphere forming capabilities, GBM 276 and GBM 612, were cultured in DMEM/F12 (1:1), 1% HyClone Antibiotic Antimycotic Solution (Thermo Scientific), Gibco B-27 serum free Supplement (Thermo Scientific 17504044), 20 ng/ml EGF, and 20 ng/ml FGF. For ligand treatment experiments, Gibco B27 serum-free supplement, minus insulin (Thermo Scientific A1895601) was used. For adherent culture, dishes were coated with Laminin (Sigma-Aldrich L2020). Coating was done in DMEM/F12 basal media for at least 2 h or overnight, and for every 1 cm² of surface area, 1 μl of Laminin was added. Drugs used in this study are as follows: Cycloheximide (Cell signaling #2112), Bafilomycin A1 (InvivoGen CAS # 88899-55-2), MG-132 (InvivoGen CAS # 133407-82-6), Cryptotanshinone (Selleckchem Catalog No. S2285), Stattic (Selleckchem Catalog No. S7024), and S3I-201 (Selleckchem Catalog No. S1155).