Nci h23 h23

Non-small cell lung cancer cells used in the experiments including NCI-H460 [H460] (ATCC^® HTB-177™, RRID: CVCL_0459), NCI-H292 [H292] (ATCC^® CRL-1848™, RRID: CVCL_0455), NCI-H23 [H23] (ATCC^® CRL-5800™, RRID: CVCL_1547), and A549 (ATCC^® CCL-185™, RRID: CVCL_0023) were obtained from the American Type Culture Collection (Manassas, VA, USA). H460, H292, and H23 cells were grown in Roswell Park Memorial Institute (RPMI) 1640 medium, and A549 cells was grown in Dulbecco’s Modified Eagle Medium (DMEM). All cell culture mediums were supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, and 100 units/mL of each of penicillin and streptomycin. Cells were placed in a humidified atmosphere of 5% carbon dioxide (CO₂) at 37 °C.

Human NSCLC cell line NCI-H23 (H23) was purchased from American Type Culture Collection (ATCC®, Manassas, VA). Human NSCLC cell line A549 was purchased from Cell Lines Service (Eppelheim, Germany). A549 cells were cultured in DMEM/F-12 with 2 mM L-glutamine and H23 in RPMI 1640 medium, both supplemented with 10% heat-inactivated FCS, 100 U/ml penicillin, and 100 μg/ml streptomycin. Under normoxic conditions (21% O₂) all cell lines were cultured at 37°C under 5% CO₂ and 98% humidity. Experiments under hypoxic conditions were performed at 37°C under 5% CO₂ and 1% O₂ in the automated Xvivo system G300CL (BioSpherix, Lacona, NY). A gas mixture of N₂ and CO₂ (Air Liquide, Paris, France) was connected to the system to hold on a constant O₂ of 1%, both in the working chamber and in the incubators. Before specific treatment cells were pre-incubated for 24 hours under normoxia or hypoxia. Cell numbers were determined by CASY® (Innovartis, Reutlingen, Germany). For cell authentication, the DNA (STR) profiling was performed using the PowerPlex® 16 HS System from Promega (Sunnyvale, CA, USA) and data were crosschecked by DNA profiles available through http://www.dsmz.de.