Figure 7. Itraconazole decreases cholesterol content in the plasma membrane. U87 cells were treated with DMSO or 5 μM itraconazole for 36 h. (A) The level of total cellular cholesterol was examined using the Cholesterol Assay Kit. The data are representative of 3 independent experiments. (B) Plasma membrane fraction was isolated by sucrose density gradient centrifugation, and the level of cholesterol on plasma membrane was examined using the Cholesterol Assay Kit. The data are representative of 4 independent experiments. (C) Cytoplasmic cholesterol distribution was examined by filipin staining. The data are representative of 3 independent experiments. (D) Cellular cholesterol distribution was determined by filipin staining coupled with LAMP1 immunofluorescence staining. The data are representative of 3 independent experiments. **P < 0.01; ns, not significant; Itra, itraconazole.