H1299

Human lung cancer cell lines (A549, H1299, PC9, H292, H1975) and human bronchial epithelial cell lines (Beas-2B) were purchased from the National Collection of Authenticated Cell Cultures (https://www.cellbank.org.cn, Shanghai, China). Cells were cultured in RPMI-1640 or F-12K medium (HyClone, Beijing, China) containing 10% fetal bovine serum (Ausbian, Australia) at 37 °C and 5% CO₂, according to manufacturer’s instructions.

Human lung adenocarcinoma cell lines A549 and H1299 were purchased from National Collection of Authenticated Cell Cultures (Shanghai, China) and cultured with DMEM (Hyclone, Logan, UT, USA) with 10% fetal bovine serum (FBS, Gibco BRL, Grand Island, NY, USA) in a thermostatic incubator containing 5% CO2. Human Jurkat T cell leukemia cells were purchased from National Collection of Authenticated Cell Cultures (Shanghai, China) and were co-cultured with H1299 cells in RPMI-1640 medium (Invitrogen, Burlington, ON, Canada) supplemented with 10% fetal bovine serum for co-culture experiments. A549 and H1299 cells were seeded in a six-well plate until the cell density reached 70–90% confluence, the transfection of plasmids was done with PolyJet DNA Transfection Reagent (SignaGen Laboratories, Gaithersburg, MD, USA) according to the manufacturer’s instructions.

All human NSCLC cell lines including A549, H1299, and H1650 were obtained from the National Collection of Authenticated Cell Cultures (Shanghai, China). 16HBE cells were from Sigma-Aldrich (City of Saint Louis, USA). Cells were cultured in DMEM supplemented with 10% FBS, 1% penicillin/streptomycin, and incubated at 37 °C in a humidified atmosphere with 5% CO₂.

BEAS-2B (human lung bronchial epithelial cells), A549, H1299, HCC827, and Calu-1 (NSCLC cells) were purchased from the National Collection of Authenticated Cell Cultures (Shanghai, China). Dulbecco’s modified Eagle medium (Thermo Fisher Scientific, Waltham, MA, USA) was used for culturing BEAS-2B cells, whereas Roswell Park Memorial Institute-1640 medium (Thermo Fisher Scientific) was used for culturing lung cancer cells. All media were supplemented with 1% penicillin–streptomycin and 10% fetal bovine serum (FBS). Cells were cultured in an incubator containing 5% CO₂ at 37 °C. All cell lines were cultured, maintained, and used in the range of 10 to 20 passages. The RS 2000 X-ray Biological Irradiator (Rad Source Technologies, Suwanee, GA, USA) was used to produce X-rays (225 kVp, 1.12 Gy/min). The Heavy Ion Medical Accelerator in Chiba (HIMAC) at the National Institute of Radiological Science was used to produce carbon ion beams (290 MeV/u, 1.22 Gy/min). The linear energy transfer (LET) of C290 at the entrance of the plateau was 13.3 keV/μm, whereas the LET in the spread-out Bragg peak was 80 keV/μm.

The human NSCLC cell lines A549, PC9, and H1299 were purchased from the Cell Bank of the Chinese Academy of Science (Shanghai, China) and cultured in RPMI‐1640 medium with 10% FBS. Human embryonic kidney 293T (HEK293T) cells were purchased from the American Type Culture Collection (Manassas, VA, USA) and maintained in Dulbecco's modified Eagle's medium containing 10% FBS. All cells were cultured at 37 °C with 5% CO₂ atmosphere and were resuscitated every 3 months and tested negative for mycoplasma contamination.
For all transfection procedures, standard protocols were followed in accordance with manufacturer's instructions using Lipofectamine 2000 (Invitrogen, Waltham, MA, USA).

Human immortalized lung epithelial cells BEAS-2B and human LUAD cell lines (HCC827, H460, H1975, A549, and H1299) purchased from Cell Bank of the Chinese Academy of Science (Shanghai, China) and grown in Roswell Park Memorial Institute (RPMI)-1640 medium (72400120, Gibco, USA) supplemented with 10% fetal bovine serum (FBS) (10091, ThermoFisher, USA) and 1% penicillin-streptomycin (15140-122, ThermoFisher, USA) in 37°C, 5% CO₂ incubator (Forma Steri-Cycle, Thermo Scientific, USA).

The lung cancer cell lines A549, SPCA1 and H1299 were obtained from the Cell Bank of the Chinese Academy of Science (Shanghai, China) and the American Type Culture Collection (Manassas, VA, USA), cultured in RPMI‐1640 (HyClone, Logan, UT, USA) or F12K (Gibco, Life Technology, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (HyClone). For hypoxia treatment, cells were cultured in the presence of 1% O₂ for 24 and 48 h or treated with different doses of CoCl₂ (100–500 μm; Sigma, St. Louis, MO, USA) for 48 h.