Figure 3
Simvastatin treatment and cholesterol depletion reversibly inhibit microglial phagocytosis. Isolated rat microglial cultures were treated with simvastatin (A-B, treatments as in Figure 1). 200 μM MEV was added concurrently for 24 h, while 1 mM hpβcd-cholesterol was added for 30 min at the end of the 24 h simvastatin treatment. At the end of treatment cells were incubated with carboxylate-modified green fluorescent latex beads (1x107/ml) for 3 h, fixed and immunolabelled for the microglia-specific antigen CD68. Phagocytosis was quantified as the proportion of cells having taken up at least 1 bead (proportion of microglia involved in phagocytosis; A) and as the mean fluorescence per cell having taken up at least 1 bead (mean uptake per active phagocyte; B). Asterisks represent P <0.05, 0.01, and 0.001 by 2-way ANOVA with Bonferroni’s post-hoc for N = 6 (A-B) independent culture preparations.