Culture media

D-glucose, fatty acids, D-fructose, culture media, and other basic reagents were obtained from Sigma-Aldrich (St.-Louis, MO, USA).

Culture media was purchased from Sigma-Aldrich. Each thioether (5–10 mg) was dissolved in DMF (50 µL) and inoculated into cultures of C. elegans. The cultures were incubated at 28 °C and 180 rpm for 72 h. Blank experiments were carried out in the absence of C. elegans. After 72 h, the fungal biomass was removed and washed with diethyl ether. The supernatant was extracted into diethyl ether (3 × 50 mL) and dichloromethane (3 × 50 mL), and the combined organic extracts were dried over anhydrous Na₂SO₄, filtered and evaporated under reduced pressure. The extracts were analysed by ¹H and ¹⁹F NMR before further purification by HPLC.

Culture media, fetal bovine serum, penicillin-streptomycin solution, glutamine, DMSO, and sulforhodamine B (SRB) were purchased from Sigma–Aldrich (St. Louis, MO, USA). Muse kits were obtained from EMD Millipore Bioscience (Billerica, MA, USA). 6-Carboxy-2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA) was from Molecular Probes (Warsaw, Poland). The antibodies against AKT and phospho-(Ser473)-AKT were from Santa Cruz Biotechnology (Heidelberg, Germany). The PARP antibody was from Cell Signaling Technology (Danvers, MA). Anti-β-actin and anti-rabbit antibodies were from Sigma–Aldrich.

CIP, potassium hydroxide (KOH), acetic anhydride, DMSO, DCM, sodium cyanoborohydride (NaBH₄), oils, surfactants, co-surfactants, pluronic 127, culture media, fluorescent dyes, and Schiff reagent were obtained from Sigma-Aldrich, Germany. All solvents used were of analytical grade.

Cell proliferation assays were performed using the CellTiter 96 AQueous One Solution Cell Proliferation Assay (MTS) (Promega, Charbonnières-les-Bains, France), as previously described^{4 (link),7 (link),87 (link)}. Briefly, cells (5 × 10³ cells per well) were plated in triplicate in 96-well plates. After 48 h, cells were incubated with IL-1β and/or TNFα at the indicated concentrations or vehicle only for 48 h with 100 µL culture media (2% charcoal-stripped FBS) (Sigma-Aldrich). Then, 20 µL MTS solution were added to all wells and cells were incubated for 2 h at 37 °C. Absorbance was then read at 490 nm using a Multiskan microplate reader (Thermo Scientific, Illkirch, France). All values were normalized to the values obtained with vehicle-treated cells to control for unwanted sources of variation.

Fresh primary hepatocytes were isolated from the male Wistar rats (250–300 g). William’s E medium was used as culture media, (Sigma, St. Louis, MO, USA) supplemented with 1 mg/mL bovine serum albumin (BSA) (PAA Laboratories, Pasching, Austria), 10 ng/mL epidermal growth factor, 0.5 mg/mL insulin, 5 nM dexamethasone, 50 ng/mL linoleic acid, 10 units/mL penicillin, and 10 mg/mL streptomycin. The hepatocyte suspension was gently mixed and pipetted onto the collagen-coated cell seeding membrane in a spot-by-spot way so as to make the cell suspension reside on the membrane evenly. The hepatocytes were then maintained either in perfusion, static, or collagen sandwich culture (as control) in the CO₂ incubator with 5% CO₂ and 95% humidity at 37 °C. For the sandwich culture, an overlay of 0.3 mg/mL collagen was added onto the cells the following day.
The animals used in the experiments were handled under the approval of the Institutional Animal Care and Use Committee (IACUC) of the National University of Singapore (NUS). No human samples or participants were used, and no other ethics approval is required.