Mice were perfused transcardially with 4% paraformaldehyde (PFA) in PBS. The perfused brains were removed and post-fixed in 4% paraformaldehyde at 4 °C. Dissected brains were post-fixed overnight with 4% PFA in PBS. To make coronal sections, the brains were cryoprotected by overnight immersion in 30% sucrose in PBS and embedded in OCT. Floating cryosections of 50 μm were permeabilized whit PBS containing 0.5% Triton-X 100 and blocked with 2% BSA and 0.3% Triton X-100 in PBS The sections were incubated overnight at 4 °C with primary antibodies and washed with PBS, incubated with Alexa 546. (1 h at room temperature) and washed with PBS.