The primary cultured GSCs SU4 and SU5 were derived from fresh surgical specimen of human GBM tissues using a method described previously with informed consent [37 (link)]. The study was approved by the Ethics Committee of the Second Affiliated Hospital of Soochow University. Cell lines were maintained in Dulbecco’s modified Eagle’s medium/F12 (Gibco, Grand Island, NY, USA) supplemented with B-27 (Gibco, Grand Island, NY, USA), 20 ng/mL recombinant human epidermal growth factor (EGF; Gibco, Grand Island, NY, USA), 20 ng/mL basic fibroblast growth factor (bFGF; Gibco, Grand Island, NY, USA) and antibiotics (100 U/mL penicillin and 100 mg/L streptomycin (HyClone, South Logan, UT, USA)).
TMZ, Nicardipine, chloroquine (CQ), rapamycin, laminine and poly-L-ornithine solutions were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Dimethyl sulfoxide was the product of MP Biomedicals, LLC (Santa Ana, CA, USA). Antibodies against β-actin, Bcl-2, Bax, p62, LC3B, AKT, p-AKT, mTOR and p-mTOR, were purchased from Cell Signaling Technology (Danvers, MA, USA). The secondary biotinylated antibodies were from Jackson ImmunoResearch Laboratories (West Grove, PA, USA).
TMZ, Nicardipine, chloroquine (CQ), rapamycin, laminine and poly-L-ornithine solutions were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Dimethyl sulfoxide was the product of MP Biomedicals, LLC (Santa Ana, CA, USA). Antibodies against β-actin, Bcl-2, Bax, p62, LC3B, AKT, p-AKT, mTOR and p-mTOR, were purchased from Cell Signaling Technology (Danvers, MA, USA). The secondary biotinylated antibodies were from Jackson ImmunoResearch Laboratories (West Grove, PA, USA).
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