Parthenolide

Murine MCT cells are a well-characterized cell model of kidney tubular epithelium suitable to study molecular mechanisms of kidney injury (Haverty et al., 1988 (link)). MCT cells were grown in RPMI 1640 (GIBCO, Grand Island, NY) supplemented with 10% decomplemented fetal bovine serum (DFBS), 2 mM glutamine, 100 U/mL penicillin, and 10 mg/ml streptomycin, in 5% CO₂ at 37°C. For experiments, cells were stimulated with 100 ng/ml human TWEAK; 0.01 to 100 mUI/ml IFNα and IFNβ (R&D Systems Inc.); the cytokine mixture made of 100 ng/ml human TWEAK, 30 ng/ml TNFα, and 30 U/ml interferon-γ (IFNγ, PeproTech) or LPS (100 ng/ml, Merck). The following chemical inhibitors were used: 10 μM PF-06700,841 tosylate salt (Sigma-Aldrich, Merck); 50 μM amlexanox; 10 μM Parthenolide, and 2.5 μM IKK16 (MedChemExpress). All the inhibitors were added to cultured cells 1 h before the stimuli. Stock solutions of the stimuli and inhibitors were made according to the specified manufacturers’ instructions. Cells were also treated with the IFNAR neutralizing antibody for 3 h before the stimuli.