Immnunohistochemistry and immunofluorescence were performed as described earlier (Patel and Hillard, 2003). Briefly, mice were anesthetized with isoflourane and transcardially perfused with 20 ml of phosphate-buffered saline (PBS) followed by 10 ml of ice-cold 4% paraformadahyde. Brains were removed and post fixed for 24 h followed by a 48 h incubation in 30% sucrose solution. Frozen sections 30 µm thick were cut on a cryostat and stored in an ethylene glycol solution at −20°C until use.
