Isolation and culture of rat HSCs and chemicals
Primary HSCs were isolated from male Sprague-Dawley rats (200-250g) as we previously described 23. Passaged HSCs were grown in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% (v/v) fetal bovine serum (FBS). Cultured HSCs were used at Passage #4-9. Human oxidized LDL was purchased from Intracel Company (Frederick, MD) and used before expiration. The presence of endotoxin in ox-LDL was < 0.1 units/ml, as measured by Limulus Amebocyte lysate assay kit (Whittaker M.A. Bioproducts, Walkersville, MD). Wnt3a was purchased from R & D Systems (Minneapolis, MN). PPARγ antagonist PD68235 was kindly provided by Pfizer (Ann Arbor, MI). Curcumin (purity > 94%) and κ-carrageenan were purchased from Sigma (St. Louis, MO). 15-deoxy-Δ12,14-prostaglandin J2 (PGJ2), a natural PPARγ agonist, was a product of Cayman Chemical Company (Ann Arbor, MI).