Mice were perfused with EM grade 4% PFA. One hemisphere of the brain was sectioned at 50 μm, coronally on a vibratome (Leica VT1000S, Leica Inc., Nussloch, Germany), and post-fixed in 2.5% glutaraldehyde for 2 h at RT. Immunocytochemistry was performed using anti-Ctip2 antibody (1:500, Thermo Fisher Scientific, Rockford, IL, USA), and biotinylated goat anti-rat IgG (1:500, Vector Laboratories, Burlingame, CA, USA). Samples were processed for prepared for EM as previously reported [14 (link)].
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