Spc a1

A549, SPC-A1 and H1299 cells were obtained from the American Type Culture Collection (Manassas, VA, USA). A549, SPC-A1 and H1299 cells were maintained at 37 °C in 5% CO₂-humidified air in RPMI 1640 medium containing 10% FBS (Sigma-Aldrich, St. Louis, MO, USA), 100 U/mL penicillin, and 100 mg/mL streptomycin/penicillin. CD44 + cells were isolated using the CD44 microbead isolation kit (Miltenyi Biotec, Auburn, CA, USA).

The human NSCLC cell lines (PC9, SPC-A1, A549, H1299, H1975 and GLC-82) and human embryonic kidney cells (HEK-293FT) were all purchased from the ATCC. PC9, A549, H1299 and H1975 cells were grown in RPMI 1640 medium. SPC-A1, GLC-82 and HEK-293FT cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) medium supplemented with 10% fetal bovine serum and 100 IU/ml penicillin/streptomycin (Sigma). All cells were grown at 37 °C in a humidified 5% CO₂ atmosphere.

Human LUAD cell lines A549 (BNCC341254) and SPC-A-1 (BNCC100939) and the endothelial cell line HUVEC (BNCC337616) used in this study were all provided by BeNa Culture Collection (China). Endothelial cells were cultivated in CM15-1 medium containing 10% fetal bovine serum (FBS) (Lonza, USA). A549 and SPC-A-1 cells were cultivated with Roswell Park Memorial Institute- (RPMI-) 1640 medium (Sigma, USA) containing 10% FBS (HyClone, USA). All cells were cultivated in a 37°C incubator with 5% CO₂. The endothelial cell line HUVECs (c-0230-5c) were purchased from Cascade Corporation (USA) and cultured in Medium 200 with a 2% low serum growth supplement (LSGS) (Thermo Fisher Scientific, USA).
Rg3 and 5-FU were acquired from Sigma (USA) and were formulated utilizing dimethyl sulfoxide (DMSO) (Sigma, USA). In cell experiments, Rg3 or 5-FU mother liquor was diluted with DMSO upon meeting the required concentration and then dripped into cells and cultured with cells for 48 h.