Type II alveolar epithelial cells were isolated as described previously [8 (link)] and cultured in Dulbecco’s modified Eagle’s medium (DMEM; GIBCO, UK) supplemented with 10% fetal bovine serum. Cells were grown on 0.1% (w/v) gelatin-coated culture ware and then cultured in a humidified incubator at 37 °C with an atmosphere of 5% CO2.
Antibodies for western blotting against ABCA1 and b-actin were from Abcam, USA. Antibody against Phospho-Akt, phospho-ERK1/2 and phospho-p38 MAPK were from Cell Signaling Technology, USA. Anti-ERK1/2 antibody, anti-p38MAPK antibody, anti-Akt antibody were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). HRP-goat-anti-rabbit IgG and HRP-goat-anti-mouse IgG were purchased from MBL (Nagoya, Japan)BrdU proliferation ELISA kit was from Roche Applied Science, Germany.
Antibodies for western blotting against ABCA1 and b-actin were from Abcam, USA. Antibody against Phospho-Akt, phospho-ERK1/2 and phospho-p38 MAPK were from Cell Signaling Technology, USA. Anti-ERK1/2 antibody, anti-p38MAPK antibody, anti-Akt antibody were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). HRP-goat-anti-rabbit IgG and HRP-goat-anti-mouse IgG were purchased from MBL (Nagoya, Japan)BrdU proliferation ELISA kit was from Roche Applied Science, Germany.
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