Rerf lc ai

Non-small cell lung cancer cell lines A549 and RERF-LC-AI were purchased from RIKEN BRC Cell Bank (Tsukuba, Ibaraki, Japan) and CRISPR/Cas9-induced NRF2 knockout in A549 cells (clone 2-11) was constructed and kindly provided by Prof Eric Kmiec (Gene Editing Institute, Christiana Care Health System, Newark, NJ, United States). All cell lines were cultured in Dulbecco’s modified Eagle’s medium (Gibco, Thermo Fisher Scientific), with 8% of Fetal Bovine Serum (Gibco, Thermo Fisher Scientific, Waltham, MA, United States) and 1% of Penicillin-Streptomycin (10,000 U/mL, Gibco, Thermo Fisher Scientific). Cells were maintained at 37 °C under humidified conditions with 5% CO2.

All cell lines were cultured at 37°C under the conditions of a humidified atmosphere and 5% CO₂. Human hepatocellular carcinoma cell line Huh7, human embryonic kidney cell line 293T and human endometrial adenocarcinoma cell line Hec1B were obtained from Japanese Collection of Research Bioresources (JCRB) Cell Bank (JCRB0403, JCRB9068 and JCRB1193). Human lung squamous-cell carcinoma cell line RERF-LC-AI cells was provided by the RIKEN BRC through the National Bio-Resource Project of MEXT, Japan (RCB0444). The human lung adenocarcinoma epithelial cell line A549, human renal cell carcinoma cell line Caki-2 and human neuroepithelioma cell line MC-IXC were obtained from the American Type Culture Collection (ATCC CCL-185, ATCC HTB-47, and CRL-2270). The Huh7-derived hepatocellular carcinoma cell line Huh7.5.1 was provided by F. Chisari. Human marrow stromal cells (HMSCs) were obtained from Cell Applications Inc. (San Diego, CA). Except for HMSC, all cell lines were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (Sigma, St. Louis, MO) supplemented with 100 U/ml penicillin, 100 μg/ml streptomycin, and 10% fetal bovine serum (FBS). HMSCs were maintained in MF-medium with 1% FBS (Toyobo, Japan). Replicon cell lines harboring HCV-RNA were maintained in DMEM containing 10% FBS and 1 mg/ml G418 (Nakalai Tesque, Kyoto, Japan).

The human NSCLC cell lines RERF-LC-AI, RERF-LC-KJ, and LC2/Ad were obtained from Riken BRC through the National Bio-Resource Project of the MEXT (Tsukuba, Japan). The A549 cells were purchased from both Riken BRC and American Type Culture Collection (Manassas, VA), while the PC–9 cell line was obtained from IBL cell bank (Gunma, Japan). The genotypes of all cell lines were identified with STR Identifier (Applied Biosystems) or PowerPlex 16 STR system (Promega). All the cell lines were maintained in culture in RPMI 1640 medium with 2mM L-glutamine (Invitrogen) supplemented with 10% FBS (Sigma-Aldrich) (A549, RERF-LC-AI, RERF-LC-KJ and PC–9) or 15% FBS (LC2/Ad) and 50 U/ml penicillin streptomycin (Sigma-Aldrich) at 37°C in a humidified atmosphere with 5% CO₂. For cell culture work, Gefitinib (Cayman, #13166), Docetaxel (Enzo Life Sciences, BML-T129), LY294002 (Cayman, #70920), PD98059 (Cayman, #10006726) and KU55933 (Tocris Bioscience, #3544) were dissolved in DMSO (Sigma-Aldrich). Gemcitabine (Tocris Bioscience, #3259), Pemetrexed (Santa Cruz, #sc–219564), Vinorelbine (Santa Cruz, #sc–216059), Caffeine (Tocris Bioscience, #2793) and N-acetyl-L-cysteine (NAC) (Sigma-Aldrich, #A9165) were dissolved in PBS (-).