Sigma, catalog #05-636, RRID: AB_309864), chicken anti-MAP2 (EMD Millipore, catalog #AB5543, RRID: AB_571049), and mouse anti–synapsin 1 (Synaptic Systems, catalog #106011, RRID: AB_2619772). After incubation with primary antibodies and subsequent PBS washes, the cultures were exposed to secondary antibodies labeled with Alexa Fluor 488, 546, or 633, as appropriate (anti-mouse and/or anti-rabbit and/or anti-chicken from Thermo Fisher Scientific) for one hour at room temperature. To visualize cell nuclei, 4′,6-diamidino-2-phenylindole (DAPI, Sigma-Aldrich, catalog #F6057) was used for nuclear staining. After staining, the samples were rinsed with PBS and mounted for analysis. Microscopic images were captured using a confocal microscope (Zeiss LSM700) for further examination and data acquisition.
