Intracellular T. cruzi load is not affected by simvastatin or benznidazole treatment before cell infection.

Endothelial-like (EA.hy926) and HUVEC cells were incubated with 5 μM simvastatin or 20 μM benznidazole 0.025% v/v final concentration. After 24 hours, the medium was replaced, and the cells were infected at a MOI of 10. After 16 hours of infection, cells were washed twice with fresh medium and the drug-free medium was changed daily. Then, after 72 hours of culture, the cells were fixed in cold methanol, and nuclei were stained with DAPI. Each panel is a representative micrograph (40X) of T. cruzi-infected, DAPI stained cells. Each experimental condition was performed in duplicate and, for each duplicate, 10 photographs were taken. In average, there were counted 20.12±3.8 cells per photograph. Infected cells averaged 5.57±1.16 and healthy cells were 14.92±3.1. Infected to uninfected cells ration was 1:5. Controls were incubated with DMSO vehicle alone. The data are expressed as the mean ± SD from three independent experiments. NS: not significant after one-way ANOVA analysis.