Ges 1 cells

The in vitro toxicity of DON and its oxidation product (3-keto-DON) were investigated using GES-1 cells and a CKK-8 test. GES-1 cells were bought from Cobioer Biosciences Co., Ltd. (Nanjing, China), and maintained under the cultivation conditions reported by Yang et al. [50 (link)]. First, 10,000 cells/well were plated into 96-well plates and allowed to proliferate to 80% confluence before cells were collected and exposed to 3-keto-DON (0.5, 1, 3, 6, 12, 24, and 48 mg/L) and DON standard (0.5, 1, 2, 4, 8, 10, and 20 mg/L) for 24 h. Following that, the spent medium was exchanged with a fresh one containing 1 mg/mL of CCK-8, which was then incubated for 4 h. Finally, cell viability was determined using an ELISA reader to measure the optical density at 450 nm. The percentage inhibition compared to control-treated cells was calculated for each compound concentration. All analyses were performed in triplicate. Statistical differences were determined using one-way ANOVA, with p < 0.05 considered to be statistically significant. IC₅₀ values were calculated using Dr Fit software version 1.042 [61 (link)] with default setting parameters.

PSA was purchased from HuBei HengLuYuan Technology Co., Ltd (HuBei, China). Sigma supplied cholesteryl chloroformate, tetrabutylammonium bromide, DOWEX 50WX2 ion-exchange resin, IR-780 iodide, and Dex. 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine polyethylene glycol-folic acid (DSPE-PEG-FA) was obtained from Nanocs (New York, NY). Pyrene was purchased from Alfa Aesar (Ward Hill, MA). Lipopolysaccharide (LPS) and Complete Freund’s adjuvant were purchased from Sigma Corporation (Cram Ridge, NJ). HyClone Laboratories Inc. (South Logan, UT) supplied penicillin–streptomycin solution (100×). Gibco (Carlsbad, CA) provided fetal bovine serum (FBS). 0.25% trypsin, DMEM culture medium, DAPI solution were supplied by Solarbio (Beijing, China). ELISA kit was purchased from MultiSciences (LiankeBio) (Hangzhou, China). RAW 264.7 and Ges-1 cells were purchased from CoBioer (Nanjing, China).