Neuronal Differentiation and CRMP2 Assessment

Fluorescence ICC was performed according to procedures previously described^{47 (link)}. The following primary antibodies were used to assess neuronal differentiation: TuJ1 (T8660, Sigma), MAP2 (CPCA-MAP2, EnCor Biotechnology, Gainesville, FL, USA), SMI312 (SMI-312R, Covance, Princeton, NJ, USA), and GFAP (MCA-5C10; EnCor Biotechnology). To evaluate CRMP2 expression and phosphorylation levels with ICC, the same p-CRMP2^T514 and CRMP2 antibodies from Cell Signaling Technologies as for western blot analysis were used. For the high-throughput high-content screening assay, neuronal cultures were co-immunostained for p-CRMP2^T514, the dendritic marker MAP2, and the nuclear protein HDAC2 (05-814, EMD Millipore, Burlington, MA, USA).