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2 protocols using streptomycin

1

Zein Protein Cytotoxicity Assay

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Zein Protein (Merc, Germany), alcalase enzyme, lecithin, Tween 80, cholesterol, ethanol, RPMI, RNase A, and yellow tetrazolium 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) was purchased from Sigma-Aldrich (St Louise, IL). Fetal bovine serum and DMSO were provided by Thermo (Waltham, MA), PBS, trypsin/EDTA, propidium iodide (PI), Triton X100, and DCFDA/H2DCFDA Cellular ROS Assay Kit were obtained from Abcam (Cambridge, UK). Penicillin, streptomycin, and Tris-HCl buffer were purchased from GeneDirex, Inc. (Gongye Rd, Taiwan). Annexin V-FITC (EXBIO, Czech), Bax, Bcl-2, caspase-3, -8, -9, and p53 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
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2

Glioblastoma Cell Culture Protocol

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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.
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