(a) ChPs were stained for C1q (red) and C4 (green). Bar
100 µm. (b) C5 siRNA treatment blocks C5 protein
deposition in ApoE-/- ChPs; (c) ChPs were stained for C3. Ig
represents lipid; (d) Serum C3 and C5. Serum C3 and C5 protein
levels were measured by ELISA. ApoE-/-(n = 6 mice), HFD ApoE4 (n=5).
(e) High resolution confocal microscopy shows
colocalization of ApoE4 (ApoE, red) and Ig (green, represents lipid) in HFD
ApoE4-KI ChPs. ApoE-/- ChPs serve as negative controls for ApoE staining;
(f) Complement regulators are expressed in ChPs. WT (n = 5
mice); ApoE-/-(n=4); ND ApoE3 (n=6); HFD ApoE3 (n=6); ND ApoE4 (n=6); HFD
ApoE4 (n=6). (g) ChP Factor H expressed between WT and ApoE-/-
mice. WT (n=5); ApoE-/-(n=4); (h) ChP factor H protein in ChPs. White arrows
indicate lipid positive areas. Data in a,b,c,e,h are representative images
from at least 3 biologically independent mouse samples. Data in d,f,g
represent means ± SEM; Two-tailed Student's t-test was applied to
d,g; one-way ANOVA with Tukey posttest was applied to f; Gene names in
Supplementary Table 3.