Rosuvastatin sml 1264

Atorvastatin (PHR-1422) and rosuvastatin (SML-1264) were obtained from Sigma Aldrich, USA. Atorvastatin and rosuvastatin were dissolved in dimethyl sulfoxide (DMSO) at a concentration of 50 mM. PD98059 (S1177, SelleckChem) was reconstituted in DMSO at a concentration of 20 mM. LY294002 (S1105, SelleckChem) was reconstituted in DMSO at a concentration of 10 mM. Lipopolysaccharide (LPS, Sigma) and mouse epidermal growth factor (EGF, Corning) were reconstituted in sterile deionized water at concentrations of 1 mg/ml and 50 μg/mL respectively.

Confluent cells were washed with FBS-free medium and treated with either 10 mM methyl-β-cyclodextrin (MβCD; M7439, Sigma-Aldrich, St. Louis, MO, USA) dissolved in FBS-free medium or FBS-free medium without MβCD for 45 min at 37 °C in 5% CO₂. After that, the medium was removed, and the cells were washed with FBS-free medium twice. The cells that were treated with MβCD were incubated in FBS-free medium supplemented with 5 μM rosuvastatin (SML1264, Sigma-Aldrich, St. Louis, MO, USA) for 24 h, while the cells that were treated with medium without MβCD were incubated with the same FBS-free medium supplemented with 0.35% DMSO (Sigma-Aldrich, St. Louis, MO, USA) for the same duration. In the initial cholesterol depletion assays, cells were not treated with MβCD and were incubated with 5 μM rosuvastatin in FBS-free medium for 24 h.