Quantifying Amyloid-Beta Deposition

Immunohistochemical analyses were performed as previously reported [56 (link),57 (link)]. Briefly, mouse brain tissues were placed in 10% neutral buffered formalin overnight. Tissues were embedded with paraffin, sectioning and placed on slides. Deparaffinized sections were stained with 4G8 antibody (#800701, BioLegend, San Diego, CA, USA) for amyloid-β (Aβ). Staining was visualized by using HRP-specific DAB substrate kit with nickel (Vector Laboratories, Burlingame, CA, USA) and mounted in DPX mounting medium (Electron Microscopy Sciences, Hatfield, PA, USA). Image files were imported to ImageJ (NIH) for quantification. Approximately 4–6 slices per animal were used and Aβ was quantified using burden threshold.