Nci h358

Lung adenocarcinoma cell lines, including NCI-H1650, NCI-H1299, LTEP-a 2, NCI-H1975, CaLu-3, A549, PG49, NCI-H358, NCI-H1299 and HEY-293T, were obtained from ATCC. The cell lines were maintained in Roswell Park Memorial Institute (RPMI) -1640 containing 10% fetal bovine serum (FBS; Invitrogen, USA).
The lung carcinoma tissues and clinical data obtained from our institute were approved by the Institutional Review Board of China (approval ID 81470137). We also subjected the cell lysates from the clinical specimens to Western blot analysis.

HeLa (CCL-2), HepG2 (HB-8065) and NCI-H358 (CRL-5807) cells were purchased from ATCC whilst Huh7 (Riken - RCB1366) were a kind gift from Prof. Samir El-Andaloussi (KI, Stockholm), all cell lines were authenticated by STR profiling and tested negative for mycoplasma contamination. Cells were maintained at 37 °C in a humidified incubator in a complete media of DMEM + Glutamax (Huh7, HeLa, HepG2) or RPMI + Glutamax (NCI-H358 cells) both supplemented with 10% foetal bovine serum.
Stable cells expressing mCherry-GAL9 were generated by knock-in at the AAVS1 locus. Cells were seeded at 2 × 10⁵ cells/well (12-well) and transfected with mCherry-GAL9 reporter:AAVS1 zinc-finger nuclease (1:9) using FuGENE HD transfection reagent (Promega) as per manufacturer’s instructions. Cells were incubated for 48 h before the addition of 1 µg/ml Puromycin to select for stably integrated cells. Stable cells were subsequently sorted into similar expression pools by flow cytometry.

Paired NSCLC and corresponding noncancerous lung tissues were obtained from 24 consecutive patients with informed consent in our department. Tissues were snap-frozen in liquid nitrogen. NSCLC cell lines A549, NCI-H23 and NCI-H358 cells, and normal GNHu27 cells were purchased from ATCC. The cell lines were maintained in RPMI-1640 medium containing 10% FBS at 37 °C in a humidified atmosphere containing 5% CO₂. Transfection was performed using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s protocol.

NSCLC cell lines, A549, and NCI-H358 (hereafter H358) were obtained from ATCC in 2014. The cells were grown in RPMI-1640 medium with 10% fetal bovine serum (FBS) and antibiotics-antimycotics (#15240-062, Invitrogen, Carlsbad, CA, USA), at 37 °C and 5% CO_2, and controlled every month for mycoplasma contamination. Recombinant human TGF-β1 was from R&D Systems (Minneapolis, MN, USA). The inhibitors EPZ5676, SGC0946, PFI-1, and SAHA were purchased from Cayman (Ann Arbor, Michigan, USA). The antibodies are listed in Additional file 1.

Human NSCLC cell lines (NCI-H460, NCI-H358 and A549) were obtained from ATCC. All cell lines were grown in RPMI medium supplemented with 10% fetal bovine serum (FBS) and antibiotics and incubated at 37 °C in a humidified atmosphere with 5% CO₂.

Human lung cancer cell lines (NCI-H358, ATCC ® CRL-5807, lung bronchioloalveolar carcinoma; Calu-3, ATCC ® HTB-55, lung adenocarcinoma; A549, ATCC ® CCL-185, lung adenocarcinoma) and IMR-90 human lung fibroblast cell lines (ATCC ® CCL-186) were obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA). IMR-90 and Calu-3 cells were cultured in Eagle’s minimal essential medium (EMEM), NCI-H358 cells in Roswell Park Memorial Institute (RPMI)-1640 medium, and A549 cells in F-12K medium supplemented with 10% fetal bovine serum (FBS, WelGENE Inc., Deagu, South Korea) and 100 U/mL penicillin-streptomycin (WelGENE Inc.) in a humidified incubator set at 37°C with 5% CO₂. Subsequently, NCI-H358 and A549 cells were cultured in EMEM to match the medium used for IMR-90 cells. We performed cytotoxicity and cell viability assays as well as real-time cell monitoring for 120 hours to observed the effects of exposure to 50 mg/L chrysotile, 50 mg/L crocidolite, and 50 mg/L amosite on IMR-90 making IMR-90 being cytostatic, and decided to use 50mg/L as our experimental concentration for all types of asbestos [17 ]. The study was approved by the institutional review board of Yeouido St. Mary’s Hospital (Korea; approval number SC19ZNSI0015) and was in accordance with the relevant legislation.