Cholesterol depletion leads to changes in lysosomal distribution within cells.
Representative panels of lysosomal distribution in control cardiomyocyte (A) and cardiomyocytes pre-treated either with 10 mM (B) or 15 mM MβCD (C), 10 mM or (D) 15 mM HγCD (E), or 15 mM MβCD followed by 0.05 mM WSC (F). MβCD treated cardiomyocytes show significant changes in lysosomal dispersion in cell cytoplasm. After drug treatment, cells were washed and incubated with T. cruzi TCTs, M.O.I. of 50 parasites per cell for 40 minutes at 37°C. After cell invasion, cells were washed, fixed and immunostained for LAMP-1 (green) DAPI (blue) and analyzed under fluorescence microscope. In comparison to untreated cardiomyocytes (A), which exhibit homogenous and well distributed LAMP-1 labeling, MβCD treated cardiomyocytes, (B) and (C), show a heterogeneous LAMP-1 labeling with lysosomes localized predominantly near cell nuclei. On the other hand, HγCD treated cardiomyocytes and cholesterol-replenished cells, (E), (F) and (D), present lysosomal distribution similar to control cells. Scale bar: 10 µm.