Glioblastoma Cell Culture Protocol

GBM cells were maintained following a previous study [35 (link)]. The GBM cell lines U-87 MG and T98G were obtained from BCRC (#60360, Taiwan) and JCRB (JCRB9041, Japan), respectively. A patient-derived GBM line P#5, was obtained according to the Taipei Medical University IRB protocol (201006011). TMZ-resistant P#5 TMZ-R cells were established from P#5 cells with long-term 50 μM TMZ exposure and provided by Professor Jian Ying Chuang (Taipei Medical University, Taiwan) [36 (link)]. Cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) with 10% fetal bovine serum (FBS, HyClone, Logan, UT, USA), 100 U/mL penicillin, and 0.1 mg/mL streptomycin (GeneDirex, Taoyuan, Taiwan). All cells were maintained in a humidified incubator with 5% CO₂ at 37 °C. LMK235 was purchased from InvivoChem. PBA, CI-994, and SW-100 were obtained from MedChemExpress. Antibodies against PARP1, MAP1LC3, and SQSTM1 were obtained from Cell Signaling Technology (Danvers, MA, USA). Antibodies against acetyl-TUBA and GAPDH were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Antibodies against SCNN1A and ACTIN were obtained from Abcam and Millipore (Burlington, MA, USA), respectively.