Rat anti lamp 2

Primary antibodies were as follows: mouse anti-a2V (Covance, Denver, PA); rabbit anti-GAPDH (Cell signaling, Danvers, MA); rat anti-F4/80, rabbit anti-LC3B, rabbit anti-LAMP-1, rat anti-LAMP-2, rabbit anti-ATG3, rabbit anti-ATG4, rabbit anti-ATG7, rabbit anti-NF-κB p65 (all from Abcam). Secondary antibodies were as follows: goat anti-rabbit IgG-FITC, -mouse IgG AF-594, -rabbit IgG AF-594 (Invitrogen), rabbit anti-rat IgG-FITC (Abcam), goat anti-rabbit, -rat, -mouse IgG-HRP (Santa Cruz Biotechnology), donkey anti-rabbit IRDye-800CW (LI-COR Bioscience, Lincoln, NE) and EnVision + dual link System-Horseradish peroxidase (HRP) (Dako). Isotype control antibodies were as follows: control mouse IgG (R&D Systems); rat IgG isotype, mouse IgG isotype and rabbit IgG isotype (Abcam).

Twelve-fourteen-month-old tg-ArcSwe mice were perfused with isotonic saline solution followed by 4 % phosphate-buffered formaldehyde (Histolab AB). The brains were frozen and cryo-sectioned sagittally to a thickness of 14 μm and then permeabilized and blocked in 0.3 % Triton X-100 in PBS containing 5 % NGS for 1 h in RT. The sections were incubated at 4 °C O/N with primary antibody diluted in 0.3 % Triton/PBS with 0.5 % NGS and thoroughly washed in 0.3 % Triton/PBS. Thereafter, sections were incubated for 1 h in RT with secondary antibody diluted in 0.3 % Triton/PBS with 0.5 % NGS before repeating the washing step. The sections were mounted on microscope glass slides with Vectashield containing DAPI (Vector). Primary antibodies used were: rabbit anti-GFAP (1:400, DakoCytomation), mouse anti-GFAP (1:500, Sigma), rat anti-LAMP-2 (1:400, Abcam) and mAb158 (5 μg/ml). Secondary antibodies used were AlexaFluor 488 against mouse, rabbit and rat or AlexaFluor 555 against mouse and rabbit (1:500, Molecular probes).