Huh7 cells were transfected with pGL4.32 [luc2P/NF-kB-RE/ Hygro] (NF-kB reporter) vector (500 ng) using Lipofectamine 2000 in OptiMEM. Renilla CMV (1 ng) was co-transfected. Cell medium was changed 4 h post-transfection and stimulated after 24 h with either sACE2/ACE, 100 ng/ml, IL-1b, 100 ng/ml, diluted in DMEM containing 1% FCS. After treatment, luciferase levels were analysed using Dual-Luciferase Reporter Assay System following manufacturer's instructions.