β cyano l alanine bca

MAC-T cells were purchased from the ATCC Corporation (Beijing, China), and maintained in dishes or plates in a humidified atmosphere of 5% CO₂ at 37 °C in a DMEM culture medium (Gibco, Grand Island, NY, USA) containing 10% fetal bovine serum (FBS, Gibco, Grand Island, NY, USA). The cells were grown to 80% confluence, and cells were passaged or treated with different compounds such as LPS (0–200 μg/mL, Solarbio, Beijing, China), β-cyano-L-Alanine (BCA, 0.5–4.0 mM, Cay-man Chemical, Ann Arbor, MI, USA), NaHS (30 µM, Sigma, St. Louis, MO, USA), or/and SB225002 (0–10 μM, Selleck, Houston, TX, USA). The cells were seeded at 1 × 10⁴ cells/well in 96-well plates or 1 × 10⁵ cells/cm³ in different dishes. Before treatment, the culture medium was changed to DMEM without FBS and phenol (Gibco) for 12 h. The MAC-T cells were treated with or without LPS, BCA, NaHS, or/and SB225002 for 24 h. The appropriate concentration gradient was selected based on literature reports [36 (link),37 (link),38 (link)] and cell viability test results, and adjusted according to the reagent situation. Each treatment was replicated at least in triplicate.