Dbtrg 05mg

Human GB cell line T98G (ATCC^® CRL-1690™) (ATCC, Manassas, VA, USA) and U87MG (ATCC^® HTB-14™) (ATCC) were cultured in DMEM; DBTRG-05MG (ATCC^® CRL-2020™) (ATCC) and GBM8901 (Bioresource Collection and Research Center, Hsinchu, Taiwan) were cultured in RPMI. In order to demonstrate the association between BC200 and MGMT, the U87MG cells were transfected to overexpress MGMT. The human MGMT open reading frame (ORF) plasmid was purchased from OriGene (Cat# MGMT (RC229131, Taipei, Taiwan) and the cells transfected according to vendor’s instructions; MGMT-overexpressing U87MG cells were then allowed to grow at 37 °C in 5% CO₂ humidified atmosphere in Dulbecco’s modified Eagle’s medium (DMEM). Media was supplemented with 10% fetal bovine serum, and streptomycin (100 μg/mL), penicillin (100 IU/mL), and at 80% confluency the cells were sub-cultured every 2–3 days. The expression of MGMT was verified by western blots.

GBM cell lines were purchased from American Type Culture Collection (ATCC, USA) and grown in a humidified, 5 % CO2 incubator at 37 °C in the complete media as described by the provider. DBTRG-05MG (ATCC, USA, #CRL-2020) cells were grown in ATCC-formulated RMPI-1640 medium (Gibco, Life Technologies, USA, #A10491-01) supplemented with 10 % foetal bovine calf serum (Gibco, Life Technologies, USA, #16000044), non-essential amino acids (Gibco, Life Technologies, USA, #11140050) and 100 U/mL penicillin, 0.1 mg/mL streptomycin (Gibco, Life Technologies, USA, #15140122) to generate complete media. M059J (ATCC, USA, #CRL-2366) cells were grown in media containing a 1:1 mixture of Dulbecco’s Modified Eagle’s Medium and Ham’s F12 Medium (DMEM-F12, ATCC, USA, #30-2006) supplemented with 10 % foetal bovine calf serum, non-essential amino acids and 100 U/mL penicillin, 0.1 mg/mL streptomycin. LN18 (ATCC, USA, #CRL-2610) and LN229 (ATCC, USA, #CRL-2611) were grown in Dulbecco’s Modified Eagle’s Medium (DMEM, ATCC, USA, #30-2002) supplemented with 5 % foetal bovine calf serum and 100 U/mL penicillin, 0.1 mg/mL streptomycin. HeLa cells were grown in a humidified, 5 % CO2 incubator at 37 °C in the complete media, Dulbecco’s Modified Eagle’s Medium (DMEM, ATCC, USA, #30-2002) supplemented with 10 % foetal bovine calf serum and 100 U/mL penicillin, 0.1 mg/mL streptomycin.

Human GBM cell lines—LN229 (CRL‐2611), U‐87 MG (HTB‐14), LN18 (CRL‐2610), A‐172 (CRL‐1620), T98G (CRL‐1690), and DBTRG‐05MG (CRL‐2020)—were purchased from ATCC. HEK293T cell line was purchased from Invitrogen (#R70007). These cells were cultured in Dulbecco's modified Eagle's medium (DMEM; 10‐013‐CV, Corning) supplemented with 10% fetal bovine serum (FBS; Gibco, 10 437 028) and 1% Antibiotic–Antimycotic Solution (30‐004‐CI, Corning) at 37 °C with 5% CO₂. None of these cell lines were listed in the database of misidentified cell lines maintained by ICLAC and NCBI Biosample. These cell lines were not authenticated in this study. All cell lines were confirmed as Mycoplasma negative before experiments. Unless otherwise indicated, cells were grown to 50% confluence.

Human glioblastoma cell lines were obtained as follows: U-251 MG, T98G, SF126, A-172, AM38, and YH-13, JCRB Cell Bank (Osaka, Japan); DBTRG-05 MG, LN-229, LN-18 and M059K, ATCC (Manassas, VA). These cell lines were maintained in RPMI1640 supplemented with 10% fetal bovine serum (FBS) and cultured at 37°C in a humidified atmosphere containing 5% CO₂.

The GBM cell lines U87, DBTRG-05MG, U118MG, and LN229 were purchased from ATCC (Manassas, VA, USA). U87 cells were grown in minimum essential medium (MEM) supplemented with 10% fetal bovine serum (FBS) at 37°C in an atmosphere of 5% CO₂. DBTRG-05MG cells were grown in RPMI-1640 supplemented with 10% FBS at 37°C in an atmosphere of 5% CO₂. U118MG and LN229 cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% FBS at 37°C in an atmosphere of 5% CO₂. The p53 status of these cell lines was confirmed using DNA sequencing analysis (Supplementary Figure S7). Cells at 70% confluency were washed with phosphate-buffered saline (PBS) and treated with TMZ (0–10 mM) and/or VPA (2.5 mM) in complete culture medium at 37°C in the dark.