Sc 13040

Immunohistochemistry was performed on 12 μm (embryonic brains) or 30 μm (postnatal brains) coronal sections (Liu et al., 2018 (link)). Briefly, sections were blocked for 30 min in TBS with 0.1% Triton X-100 and 5% donkey serum. For double staining, sections were incubated simultaneously with primary antibodies from different species, and secondary antibodies were used sequentially. Primary antibodies were incubated for 24 h at 4°C. We used rabbit anti-calretinin (CR; 1:3,000, AB5054, Millipore, Burlington, MA, USA), chicken anti-GFP (1:2,000, GFP-1020, Aves Labs), rabbit anti-PV (1:2,000, PV25, Swant), rabbit anti-NKX2-1 (1:500, sc-13040, Santa Cruz Biotechnology, Dallas, TX, USA), rabbit anti-NPY (1:500, 22940, Incstar), goat anti-SST (1:500, sc-7819, Santa Cruz Biotechnology, Dallas, TX, USA) and goat anti-SP8 (1:2,000, sc-104661, Santa Cruz Biotechnology, Dallas, TX, USA). Secondary antibodies (1:400, Jackson Immuno Research) were incubated for 2 h at room temperature (RT), rinsed three times in TBS, and then incubated with DAPI (4′,6-diamidino-2-phenylindole, 1:5,000) for 3 min.