Nci h358

A549 (10185), NCI-H358 (25807) and Calu-1 (30054) lung cancer cells were obtained from Korean Cell Line Bank (Seoul, South Korea). Dulbecco’s modified Eagle’s medium (DMEM) and Roswell Park Memorial Institute (RPMI) 1640 with 10% fetal bovine serum (FBS) and antibiotics were used for cell culture. For transient transfection, mammalian expression vectors were transfected by using Polyfect (Qiagen, Hilden, Germany) in HEK293T, A549, Calu-1 and NCI-H358 cells. After transfection, cells were further incubated for 48 h to allow stabilization and protein expression.

HDFs (PromoCell GmbH, Heidelberg, Germany) and HeLa cells (ATCC, Manassas, VA, USA) were cultured in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS, Gibco, Grand Island, NY, USA) and 0.1% antibiotics (Gibco) at 37℃ in a 5% CO₂ incubator (APM-30D; ASTEC, Fukuoka, Japan) [33 (link)]. Oxygen levels of 21%, as the normoxic condition, and 1%, as the hypoxic condition, were used for culturing HDFs from passage 4 to passage 6 [33 (link)]. When the cell confluency of all cells reached 90%, the cells were passaged using 0.25% Trypsin-EDTA (Gibco).
For the proliferation assay, 2 × 10⁵ HDFs at passage 6 were cultured in a 100-mm culture plate for 5 days, and cell numbers were measured using trypan blue 0.5% solution staining (Biowest, Riverside, MO, USA). Other cervical cancer cells (CaSki, ME-180, and SiHa cells), melanoma cells (A375SM), breast cancer cells (MCF-7), and lung cancer cells (NCI-H358) were purchased from the Korea Cell Line Bank (Seoul, Korea). CaSki, ME-180, A375SM, MCF-7, and NCI-H358 were cultured in RPMI1640 (Gibco) supplemented with 10% FBS (Gibco), 1% L-glutamine (Gibco), 25 mM HEPES (Gibco), 25 mM NaHCO₃ (Gibco), and 0.1% antibiotics (Gibco). SiHa cells were cultured in DMEM supplemented with 10% FBS (Gibco) and 0.1% antibiotics (Gibco).

Human cervical cancer HeLa cells, as well as non-small cell lung cancer NCI-H23, NCI-H1703, NCI-H358, and Calu-3 cells, were purchased from the Korean Cell Line Bank (Seoul, Korea) or KRIBB Cell Line Bank (Daejeon, Korea). The lung cancer cell lines YL01, YL03, YL05 (EGFR exon19del), and YL08 (EGFR wild-type/ALK mutation-positive), each derived from a different patient, were provided by the Yonsei University College of Medicine, Seoul, Korea. Patient characteristics and treatments were previously described^{19 (link)}. Cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) or RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) and penicillin/streptomycin (Invitrogen, Carlsbad, CA, USA). Cells were transfected with plasmids using Turbofect (Thermo Scientific, Rockford, IL, USA) and with siRNAs (20–40 nM) by electroporation (Neon, Invitrogen) according to the manufacturer’s instructions. The siRNAs used in this study were purchased from Bioneer Corporation (Daejeon, Korea). The target sequences were as follows: siJNK1 #1, 5′-CUGGUAUGAUCCUUCUGAA-3′; siJNK1 #2, 5′-GUCACACCUGGAAACCUGA-3′; and siScrambled, 5′-CCUACGCCACCAAUUUCGU-3′.

In this experiment, we used lung cancer cell lines (HCC827, NCI–H358, NCI–H522, NCI– H1299, NCI– H460, and NCI–H226) and HeLa cell line. We purchased the HCC827, NCI–H358, NCI–H522, NCI– H1299, NCI– H460, and HeLa from the Korea Cell Line Bank, and we purchased the NCI–H226 from the American Type Culture Collection. Lung cancer cells (HCC827, NCI–H358, NCI–H522, NCI– H1299, NCI– H460, and NCI–H226) and HeLa cells were maintained in an RPMI 1640 medium (GIBCO BRL, Rockville, MD, USA) with 10% fetal bovine serum and antibiotics (100 units/mL penicillin and 100 mg/mL streptomycin).