Hepg2

The dried algal biomass of S. maxima was supplied by the Hydrobiology Lab of the Water Pollution Research Department at the National Research Centre in Giza, Egypt. Tetraethyl orthosilicate (TEOS) and Tween 80 were acquired from Scientific Fischer Co. (USA). The cancer cell lines MCF-7 and HepG2 were purchased from the American Type Culture Collection (ATCC), located in Virginia, (USA). Tissue culture media was obtained from Invitrogen-Life Technologies. MTT was purchased from Acros Organics™ Thermo Fisher Scientific, USA. DOX was obtained from the drugstore in Egypt. Female CD1 Swiss albino mice were provided by the animal facility at the National Research Centr in Giza, Egypt.

The human hepatocarcinoma cell line HepG2 was purchased from the American Type Culture Collection (ATCC HB-8065). Dulbecco’s Modified Eagle Medium (DMEM-GlutamaxTM) with high glucose (4.5 g/L), Phosphate-Buffered Saline (PBS), Newborn Calf Serum (NBCS), penicillin, streptomycin, and trypsin-EDTA 0.5% were obtained from Gibco (Thermo Fisher Scientific, Waltham, MA, USA). Dimethyl sulfoxide (DMSO) and methanol were acquired from Fisher Scientific (Madrid, Spain). The standards of AZX (purity ≥ 98%, MW 403.39 g/mol), DON (purity ≥ 98%, 296.32 MW g/mol), OTA (purity ≥ 97%, MW 403.81 g/mol), T2 (purity ≥ 98%, MW 466.52 g/mol), thiazolyl blue tetrazolium bromide (MTT), and the resazurin sodium salt were supplied by Sigma-Aldrich (St Louis, MO, USA). All the reagents and cell culture components were of standard laboratory grade.
The fungicide and the mycotoxins were prepared as stock solutions in DMSO and methanol, respectively. These solutions were stored at −20 °C until use. Working concentrations were then prepared in a DMEM-supplement medium with a maximum solvent concentration of 0.1% in the test solutions. Negative controls containing the appropriate amounts of solvents were included in every experiment.

The human melanoma cell lines A375, SK-MEL-28, human lung cell line A549, human colorectal cancer cell lines HT29, RKO, human hepatocellular carcinoma cell line Huh-7, HepG2, human fibrosarcoma cell line HT1080 and human embryonic kidney cell line HEK293T were obtained from the American Type Culture Collection (ATCC), and human melanoma cell line SK-MEL-30 was given from Dr. Jun Wan’ laboratory in Shenzhen PKU-HKUST Medical Centre in China. All cell lines were cultured in DMEM medium (Gibco, C11995500BT) supplemented with 10% fetal bovine serum (FBS) (Excell, FSP100) at 37 °C incubator with 5% CO₂.
All cells were cultured in a 10 cm plate and subcultured into 96-well- or 6-well plates for cell death, cell viability and lipid-peroxidation measurements. The cells were treated with reagents including the ferroptosis inducers RSL3 (Selleck, S8155) and erastin (Selleck, S7242), ferroptosis inhibitors Ferrostatin-1 (Fer-1) (Selleck, S7243), apoptosis inhibitor Z-VAD-FMK (Z-V) (Selleck, S7023), necrosis inhibitor Necrostatin-1 (Nec-1) (Selleck, S8037), autophagy inhibitor Bafilomycin A1 (Baf-A1) (Selleck, S1413), SLC25A1 inhibitor BTA (Macklin, B801861), ACLY inhibitor SB204990 (Targetmol, T16861), dimethyl Citrate (MCE, HY-N9542), and sodium acetate (Targetmol, T23377).