HepG2 cells were obtained from Dr Xin Chen's laboratory at the University of California, San Francisco. Sodium oleate was obtained from Sigma-Aldrich and was dissolved in Dulbecco's modified Eagle medium (DMEM) with 1% fatty acid free bovine serum albumin (BSA) (Sigma). Oleate treatment of HepG2 cells was carried out as previously described with minor revision.10 (link)
20 (link) Specifically, HepG2 cells were plated in four-well chamber slides with DMEM medium supplemented with 10% fetal bovine serum (Invitrogen). After 24 h, HepG2 cells were treated with either control medium (DMEM supplemented with 1% fatty acid free BSA) or medium containing oleate (0.5 mM). The cells were cultured for another 24 h, then lipid accumulation and miR-21 expression were determined by Nile Red Staining (Sigma-Aldrich) and qRT-PCR, respectively (see online supplementary materials and methods for details).
20 (link) Specifically, HepG2 cells were plated in four-well chamber slides with DMEM medium supplemented with 10% fetal bovine serum (Invitrogen). After 24 h, HepG2 cells were treated with either control medium (DMEM supplemented with 1% fatty acid free BSA) or medium containing oleate (0.5 mM). The cells were cultured for another 24 h, then lipid accumulation and miR-21 expression were determined by Nile Red Staining (Sigma-Aldrich) and qRT-PCR, respectively (see online supplementary materials and methods for details).
