Brain sections and astrocytes cultured on coverslips were immunostained for GFAP or neurocan as previously described [19 (link),29 (link),30 (link)]. The sections or cells were permeabilized with 0.1% Triton X-100 for 1 h at 37°C after formaldehyde fixation. Mouse anti-GFAP (1:200; Epitomics) or goat antineurocan (1:100, Santa Cruz Biotechnology, Dallas, TX, USA) primary antibody was applied in 5% bovine serum albumin (BSA) overnight at 4°C. After repeated washes, the cells or brain sections were incubated with FITC-conjugated secondary antibody in 5% BSA for 1 h at 37°C. The coverslips or tissue sections were mounted on glass microscope slides and photographed under fluorescent microscopy (Olympus BX51, Olympus Optical, Tokyo, Japan).