Figure 5

Atorvastatin promotes endothelial expression of Akt: A. Western blot. HLEC were pre-treated with different concentrations of atorvastatin (0.05, 1 or 5 microM) for 24 h and then cultured or co-cultured with PRBC (at 5% of hematocrit and 50% of parasitemia) or RBC (at 5% of hematocrit) for 4 h. Cells were harvested, washed, protein extracts separated by SDS-PAGE, transferred to nitrocellulose and immunoblotted with anti-Akt and anti-tubulin antibodies, the latter as internal control. Molecular mass of the corresponding proteins is shown. The Western blot data were then quantitatively analysed by densitometry imaging (ratio Akt/Tubulin). Similar results were obtained in three independent experiments. B. Confocal microscopy. HLEC were pre-treated with different concentrations of atorvastatin (0.05, 1 or 5 microM) for 24 h and then exposed to PRBC (at 5% of hematocrit and 50% of parasitemia) or RBC (at 5% of hematocrit) for 4 h. Fixed cells were stained with anti-Akt antibody and analysed by confocal microscopy. Similar results were obtained in 3 independent experiments. (Bar: 20 μm).