Immortalized Hepatic Stellate Cell Culture

The immortalized human hepatic stellate cell line LX-2 was purchased from Xiang Ya Central Experiment Laboratory (China) and cultured in Dulbecco's Modified Eagle Medium (DMEM, Gibco, USA) supplemented with 10% fetal bovine serum (FBS, Invitrogen, USA) in a humidified incubator at 37°C with 5% CO₂. SEA of S. japonicum was obtained from Jiangsu Institute of Parasitic Diseases (China). Primary antibodies for mouse double minute protein 2 (Mdm2), total-Akt (T-Akt) and p53 were purchased from Santa Cruz Biotechnology (USA). Primary antibodies for caspase-3 and phospho-Akt (Ser⁴⁷³, p-Akt) were purchased from Cell Signaling Technology (USA). The primary antibody for glyceraldehyde phosphate dehydrogenase (GAPDH) was provided by Goodhere (China). All of the secondary antibodies were obtained from Santa Cruz Biotechnology (USA). In addition, GW501516 (Santa Cruz, USA), which is a potent PPARβ/δ agonist, was also used to activate Akt signaling [15] (link).