Intracellular cholesterol levels and phagocytosis in macrophages in presence of simvastatin.

<p>(A) Representative images of simvastatin ± mevalonate-treated macrophages overnight. Cells were washed and then stained with filipin and (B) fluorescent intensity (arbitrary units) per cell was quantified by Laser Scanning Microscope (LSM) software. Data is shown as intensity from 50-100 cells/group (Original magnification x100). (C) Macrophage cholesterol levels were measured using filipin staining 1 hour after <i>L. monocytogenes</i> infection. (D) Cholesterol content was measured in simvastatin-treated macrophage cell lysates following lipid extraction. (E) Macrophages were treated with either simvastatin ± mevalonate for 24 hours or ± methyl-β-cyclodextrin (MβCD) or with cholesterol for 2 hours, and then incubated with latex beads at MOI=10 to measure phagocytosis (Original magnification x100). Cells were then analyzed for number of internalized beads in each setting. (F) Uptake of GFP-expressing Listeria was measured in simvastatin-treated macrophages at 90 minutes post-infection. (G) Extracellular growth of <i>L. monocytogenes</i> was determined in tryptic soy broth supplemented with indicated concentrations of simvastatin. Results are shown as mean ± SEM of triplicates and are representative of two or three independent experiments, * <i>p</i> < 0.05, ** <i>p</i> < 0.01 versus control.