Transgenic mice, expressing hAPP, line J9 were selected for this study.
Western Blot analysis of mouse brains were probed by rabbit polyclonal anti-RAP, 1:20,000 and anti-LAP, 1:1000, gifts from Dr.Robert Orlando (Univ. New Mexico, Alburquerque, NM); monoclonal anti-LDL-R, 1:1000 (Calbiochem) and anti-hAPP, 8E5, 1:10,000 (gift from Elan Pharmaceuticals).
Amyloid deposits were immunohistochemically stained with mouse monoclonal antibody 3D6, 1:600 (gift of Elan Pharmaceuticals). For double staining the brain sections were stained with rabbit polyclonal anti-amyloid-beta R1280 1:500 (gift from Dr. Dennis Selkoe, Brigham and Woman's Hospital, Boston, MA) and either anti-phosphorylated tau AT8, 1:100 (Innogenetics) or anti-phosphorylated neurofilaments SMI312, 1:1000 (Sternberger Monoclonals, Baltimore, MD).
Sandwich ELISA assays were used to detect Abeta(1-40) and Abeta(1-42) (Biosource).
Neurodegeneration was determined by staining brain sections with monoclonal anti-GFAP 1:500 (Chemicon) or anti-MAP2 1:10 (Chemicon).