Immunohistochemistry for Brain Sections

IHC was performed as previously described (Ulmke et al., 2021 (link)). Briefly, the antigen retrieval was performed by incubating the brain sections in 0.01 M sodium citrate buffer for 60 min at 70°C, followed by cool-down for 20 min at room temperature. The sections for IHC were then incubated overnight with primary antibody at 4°C after blocking with 5% normal sera of the appropriate species. Incubation with primary antibodies was followed by a 1 h incubation at room temperature with the appropriate A488-, A594-, A555- or A647-labeled (Alexa series, Invitrogen, 1:400) secondary goat or donkey antibodies. Sections were later counterstained with Vectashield mounting medium containing DAPI (Vector laboratories) to label nuclei.