Fig. 4

Effects of complement depletion on leukocyte recruitment into the brain parenchyma after i.c.v. LPS injection. a WT C57BL/6J mice were treated i.c.v. with LPS, and C3 levels in the brain (without dilution, cleared from blood proteins) and plasma (diluted in 1:100) were determined by Western blotting. b WT C57BL/6J mice received i.p. injections of CVF (10 μg/mouse) or PBS. After 4 h, WT C57BL/6J mice (n = 4) received i.c.v. LPS injection, and after 4 h, plasma and brain samples were then collected for C3 Western blotting analysis. At 4 h after CVF injection, WT C57BL/6J mice (n = 4) received i.c.v. LPS injection, and plasma samples were collected for C3a ELISA assay (c). At 4 h after i.p. injection of CVF to deplete complement from the blood circulation, mice received i.c.v. LPS injection. Then, after 4 h, rolling and adhesion leukocytes were quantified using intravital microscopy (d). e P-selectin and E-selectin mRNA expressions were measured using real-time PCR. f VCAM-1 and E-selectin protein levels in the brain were examined by Western blotting (n = 4). Images are representative of four experiments. The ratio of VCAM-1 and E-selectin to β-actin was determined through densitometry. The results are shown as the means ± SEM. *P < 0.05, **P < 0.01; n ≥ 4 for all groups