Immunostaining of Brain Sections

The brain sections were prepared as described previously (22 (link), 55 (link)). In brief, the brain tissues were sagittally sectioned 30 μm thick with a Cryostat (CM3000; Leica). Immunostaining was performed as described previously (22 (link), 34 (link), 57 (link)). Briefly, after blocking nonspecific protein binding, the sections were then incubated with a monoclonal antibody 6E10 (catalog no.: SIG-39320; 1:2000 dilution; Covance) and monoclonal antibody against GFAP (catalog no.: SMI-22R; 1:5000 dilution; Covance). Then sections were incubated with biotinylated secondary antibody of horse antimouse IgG. Following washing, the Vectastain kit (Vector Laboratories) was applied. Samples were visualized using 3,3′-diaminobenzidine as a substrate (Vector Laboratories). The sections were processed deleting primary antibody as negative controls and counterstained with hematoxylin (Sigma–Aldrich).