Amphotericin b

The LS174T, a well-differentiated human colonic goblet cell line [19 (link)], was purchased from Sigma-Aldrich (product number 87060401-1VL Saint Quentin Fallavier, France). Cells were routinely grown as a monolayer in 75 cm² plastic flasks and cultured at 37 °C in a humid environment containing 5% CO₂ in MEM medium supplemented with 2 mM L-Glutamine (Dutscher, Brumath, France); 10% heat-inactivated fetal bovine serum (FBS); 1% non-essential amino acids (Dutsche, Brumath, France); and an antibiotics cocktail containing 100 U/mL penicillin, and 100 mg/mL streptomycin and amphotericin B (Dutscher, Brumath, France). Cells were seeded in six-well plates with 4 × 10⁵ cells/well. When cells reached 80% confluence, they were treated with different stimuli. PAL, DHA, EPA, LNA, OA, AA, and LA were dissolved in 100% ethanol at a 1 mM concentration, flushed with nitrogen, and stored at −20 °C until use. Before treatment, fatty acids were placed at room temperature, except for PAL, which was heated in a 40 °C water bath. Fatty acids were added to 1 mL of MEM containing FA-free BSA to be combined at a 4:1 ratio. The specific treatment concentrations and incubation times are shown in the figure legends. For the control, cells were treated with the same quantity of ethanol and MEM-BSA concentrations as cells treated with FA.