Si smile

Human hepatoma cell lines (HepG2 and Huh7 cells) and a mouse immortalized hepatocyte cell line (AML12) were obtained from the ATCC and cultured as previously described [20 (link)]. SuperFect (QIAGEN, Hilden, Germany) or polyethylenimine (Polysciences, Inc., Warrington, PA, USA) were used for transient transfections according to each manufacturer’s recommendations. Small interfering RNAs (si-Con and si-SMILE) (QIAGEN) were transfected into HepG2 cells using Lipofectamine RNAi MAX (Thermo Fisher Scientific, Waltham, MA, USA). The luciferase assay was carried out as described previously [31 (link)]. In brief, HepG2 cells were transfected with indicated reporter plasmids together with vectors expressing SMILE, ALK3-CA, or HFE2 followed by treatment with BMP-6 or EGCG for 12 h.

Small interfering RNAs against control (si-Con) and human SMILE (si-SMILE) were purchased from QIAGEN (Cat. No. 1027416). HepG2 cells were transfected with si-Con and si-SMILE using Lipofectamine RNAi MAX (Thermo Fisher Scientific) according to the manufacturer’s instruction.