Immunohistochemical Staining for Neurodegenerative Markers

Immunohistochemical staining was performed according to a previously described method (Gu et al. 2015 (link)). The sections were reacted with specific primary antibodies for Aβ (1:300; Abcam, Inc., Cambridge, MA, USA), glial fibrillary acidic protein (GFAP) (1:300; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), ionize calcium-binding adapter molecule 1 (IBA-1) (1:300; Abcam, Inc., Cambridge, MA, USA), inducible nitric oxide synthase (iNOS) (1:300; Novus Biologicals, Inc., Littleton) and cyclooxygenase-2 (COX-2) (1:300; Cell Signaling Technology, Inc., Beverly, MA, USA). Tissues were then incubated with the corresponding conjugated secondary antibodies: goat anti-rabbit, goat anti-mouse, or donkey anti-goat IgG-horseradish peroxidase (HRP) (1:5000; Santa Cruz Biotechnology Inc. Santa Cruz, CA, USA). The sections were evaluated under a light microscope (Microscope Axio Imager.A2, Carl Zeiss, Oberkochen, Germany) (×50 or ×200 or ×400).