Map1lc3

The main primary antibodies used in this study were specific for MAP1LC3 (Cell Signaling Technology, Beverly, MA, USA; Cat 2775), ATG5 (Novus Biologicals, Littleton, CO, USA; Cat NB110-53818), SQSTM1/p62 (Santa Cruz Biotechnology Inc., Heidelberg, Germany; Cat sc-25575), DAPDH (Cell Signaling Technology; Cat 2118), IFNGR1 (Bioss Biotechnology Co., LTD., Beijing, China; Cat bs-1463R), IFNGR2 (Bioss Biotechnology Co., LTD.; Cat bs-2710R), LC3B (Bioss Biotechnology Co., LTD; Cat bs-4843R) and SQSTM1 (Bioss Biotechnology Co., LTD; Cat bs-2951R). 3-Methyladenine and rapamycin were purchased from Santa Cruz Biotechnology Inc. 4′,6-diamidino-2-phenylindole (C1005) and pifithrin-α were purchased from Beyotime (Beyotime Institute of Biotechnology, Shanghai, China). HRP-conjugated goat anti-rabbit secondary antibodies were purchased from Proteintech (Proteintech Group, Inc., Chicago, IL, USA). Bovine interferon was purchased from the Kingfisher Group (Kingfisher Biotech, Inc., Saint Paul, MN, USA).

The following primary antibodies were used for western blotting: protein kinase B (Akt) (#9272), p-Akt (#4060S), p70 S6 kinase (p70S6K) (#9202), p-p70S6K (#9205), S6 ribosomal protein S6 (S6) (#2217), p-S6 (#4858S), 4E-binding protein 1 (4E-BP1) (#9452), p-4E-BP1 (#9459), and MAP1LC3 (Microtubule-associated protein 1 light chain 3 (LC3) (4108; all from Cell Signaling Technology, Danvers, MA, USA), Muscle Atrophy F-box (MAFbx) (sc-33782; Santa Cruz), Muscle RING-Finger Protein (MuRF1) (sc-32920; Santa Cruz), p62 (SQSTM1) (PM045, MBL), ubiquitin (sc-166553; Santa Cruz) AMP-activated protein kinase (AMPK) (#2532; Cell Signaling Technology), p-AMPK (#2531; Cell Signaling Technology), PGC-1α (516557; Millipore) and oxidative phosphorylation (OXPHOS) (ab110413; Abcam, Cambridge, UK).

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.