Neurons were then incubated with secondary antibody (Table 2) prepared in ice-cold blocking/permeabilization buffer for two hours at room temperature, followed by two washes with PBSCa2+-Mg2+. Nuclei were counterstained with 0.5 μg/ml DAPI (Sigma, #D9542) for 10 min and then neurons were washed with PBS three additional times.
