Quantification of ApoE and Inflammation Markers

ApoE was measured by ELISA in brain endothelial cell–enriched lysates (1:8 dilution), cortical lysates (1:160 dilution), and plasma (1:20 000 dilution) as described in previous studies^{11 (link),28 (link)} using anti-apoE (1:2000; Millipore) and biotinylated anti-apoE (1:5000; Meridian) for capture and detection antibodies, respectively. Cortical lysates were analyzed for fibrinogen (1:3 dilution), IgG (1:3 dilution), and albumin (1:100 dilution) with commercial ELISA kits (Immunology Consultants) with 2 minor modifications to the manufacturer’s protocol; sample incubation time was increased to 2 hours at 37 °C, and secondary antibody was increased to 1 hour at 37 °C, and standards were buffer matched. Cortical samples were normalized to protein concentration, and isolated brain endothelial cell samples were normalized to actin quantified by the Western blot. Plasma levels (1:10 dilution) of cardiovascular dysfunction were measured using MILLIPLEX MAP CVD magnetic bead panel according to the manufacturer’s protocol.