Neural cultures were fixed with 4% paraformaldehyde in PBS (Invitrogen) for 45 min and stained with mouse monoclonal antibody to beta III-tubulin (Millipore, 1:500), and either rabbit polyclonal antibody to s100β (Abcam, Cambridge, MA, USA 1:250) or synapsin1 (Millipore, 1:500). All primary antibodies were diluted in 0.1% bovine serum albumin in PBS. Appropriate donkey anti-mouse and donkey anti-rabbit alexa fluor 488 and 594 (Invitrogen, 1:600) secondary antibodies were used for visualization.
