DCX Immunohistochemistry Protocol

In the same manner, as described below, immunohistochemistry for DCX was performed (Park et al., 2019 (link)). The sections were treated with PBS for 10 min and then treated again with 1% H₂O₂ for 20 min. The sections were treated with goat anti-DCX antibody (1:1,000; Oncogene Research Product, Cambridge, UK) for 2 hr. The sections were treated with the biotinylated goat secondary antibody (1:500; Vector Laboratories, Burlingame, CA, USA) for another 1 hr, washed, and treated with ABC complex (1:100; Vector Laboratories). Using the 0.03% diaminobenzidine, labeling was visualized and the sections were mounted onto gelatin-coated slides. After air drying the slides at room temperature overnight, the coverslips were mounted using Permount (Thermo Fisher Scientific Inc., Waltham, MA, USA).