Figure 5.

Modulation of cellular cholesterol regulates Gc effects. HeLa cells were transiently transfected with MMTV-Luc reporter gene and GR then transferred to 10 or 50% serum overnight. Cells were treated with Genistein (as indicated) for 4 h, and then treated with 100 nM Dex for a further 4 h before lysis and luciferase assay (A). Cells were also cultured in either normal serum, charcoal stripped serum or lipoprotein-deficient serum (B and C), and then treated with 100 nM Dex for 16 h before luciferase assay. Cells were cultured in 10% serum with 10 μg/ml cholesterol (CHOL) complexed with 0.5% methyl-β-cyclodextrin (MCD) or with 0.5% MCD alone (D). Cells transfected with MMTV-Luc and GR, cultured in 10 or 50% serum were treated with Simvastatin (as indicated) together with 100 nM Dex for 16 h before luciferase assay (E). Cells were cultured in 50% serum, together with cholesterol complexed with 0.5% MCD, 0.5%, or 5% MCD alone, then treated with 100 nM Dex for 16 h before luciferase assay (F). Experiments were carried out in triplicate and repeated three times. Graphs show mean±s.e.m. *P<0.05 and **P<0.01.