H1299

The human LUAD cell lines A549 and H1299 were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA) and authenticated by short tandem repeat (STR) profiling. The HEK293T cell line was acquired from the Integrated Hospital of Traditional Chinese Medicine (TCM-Integrated Hospital) of Southern Medical University (Guangzhou, P.R. China). These cell lines were analyzed for mycoplasma and were verified to be mycoplasma-free. A549 and H1299 cells were cultured in RPMI 1640 medium (Biological Industries, Bet Haemek, Israel) supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich, St. Louis, MO, USA). HEK293T cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Biological Industries) supplemented with 10% FBS. All cells were incubated in a humidified incubator at 37°C with 5% CO₂. The AKT1 inhibitor MK-2206 2HCl and the MAPK3/1 inhibitor SCH772984 were purchased from Selleck Chemicals (Houston, TX, USA). For inhibitor treatment, 5 mmol/L MK-2206 2HCl and/or 5 mmol/L SCH772984, or dimethyl sulfoxide (DMSO; Sigma-Aldrich) alone for control, were freshly added to the cell culture every 24 h.

The NSCLC cell lines, A549 (cat no. SCSP-503) and H1299 (cat no. SCSP-589), and BEAS-2B (cat no. KCB200922YJ) were purchased from the Cell bank of the Chinese Academy of Sciences. The A549 and H1299 cells were cultured in RPMI-1640 medium (cat no. 01-100-1ACS; Biological Industries) containing 10% fetal bovine serum (FBS) (cat no. 04-001-1A; Biological Industries), 100 U/ml penicillin, and 100 mg/ml streptomycin. BEAS-2B cells were cultured in the serum-free medium for BEAS-2B cells purchased from the Cell bank of the Chinese Academy of Sciences (cat no. KCB M006). All the cell lines were cultured in a humidified chamber maintained at 37°C and 5% CO₂. The stock solution of CVB-D (100 mmol/l; cat. no. C117989; Shanghai Aladdin Biochemical Technology Co., Ltd.) was prepared in methanol.