Fig. 1. Flowchart of experimental protocol. Huh7.5 cells and human primary hepatocytes were treated with 2 mM free fatty acids (FFAs) for 3 h, in presence or absence of 200 pM TNFα administrated for 3 h in co-stimulation with 2 mM FFAs. In some experiments, genistein and 17β-estradiol were given for 30 min before FFAs administration, in presence or absence of wortmannin, MCC950, dorsomorphin, fulvestrant, G15 and TLR4-IN-C34, which were administrated for 30 min before genistein and 17β-estradiol. After the exposure to the above agents, specific assays were performed. MTT: 1% 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide; JC: 5,51,6,61-tetrachloro-1,11,3,31 tetraethylbenzimidazolyl carbocyanine iodide; DCFDA: 2,7-dichlorodihydrofluorescein diacetate; wortmannin: PI3K inhibitor; MCC950: NLRP3 inflammasome inhibitor; dorsomorphin: AMPK inhibitor; fulvestrant: estrogenic receptors inhibitor; G15: G protein-coupled estrogen receptor inhibitor; TLR4-IN-C34: toll like receptor 4 inhibitor.