Immunohistochemistry was performed on paraffin-embedded human embryonic cerebellum tissue. To expose target proteins, heat-induced epitope retrieval by 1mM EDTA in 10mM Tris buffer (pH8.5) at 120°C for 3 min. Following antigen retrieval, tissues were probed with a NEGR1 monoclonal antibody (Product # MA5-27279) at a dilution of 1:150.