Neuronal nuclei (neun)

Mouse hemibrains were fixed in 4% PFA for 24 h, cut into 10
μm cryosections, mounted onto microscope slides, and immunostained as
previously described [44 (link)]. Brain
sections were incubated with anti-Aβx-42 (1:200, Covance),
anti-Aβ 6E10 (1:1000, Covance), anti-pGlu3 IgG2b (1μg/mL;
Vivoryon Therapeutics) and NeuN (1:200, Serotec) mouse monoclonal
antibodies, or Aβ40 (1:200, Covance), Iba-1 (1:200, Wako) and GFAP
(1:1000, DAKO) rabbit polyclonal antibodies, DAPI or CD68 (1:250, Serotec)
rat polyclonal antibody overnight at 4°C. Sections were washed in TBS
and then incubated with biotinylated secondary antibodies and developed
using Vector ELITE ABC kits (Vector Laboratories) and 3,3-diaminobenzidine
(Sigma-Aldrich) or immunofluorescent-labeled secondary antibodies and
cover-slipped with mounting media (Vector).

Hemibrains were fixed in 4% PFA for 24 h and immunohistochemistry was performed as described previously (5 (link)). Fixed, frozen sections were incubated with anti-Aβx-42 (1:200, Covance), 3D6 (Aβ1–5) (1:1000, Elan), 6E10 (1:1000, Covance), MAP-2 (1:500, Millipore) and NeuN (1:200, Serotec) mouse monoclonal antibodies, or Aβ40 (1:200, Covance), Iba-1 (1:200, Wako) and GFAP (1:1000, DAKO) rabbit polyclonal antibodies, or CD68 (1:250, Serotec) rat polyclonal antibody overnight at 4°C. After washing with TBS, sections were incubated with biotinylated secondary antibodies and developed using Vector ELITE ABC kits (Vector Laboratories) and 3,3-diaminobenzidine (Sigma-Aldrich) or immunofluorescent-labeled secondary antibodies and cover-slipped with mounting media (Vector). Thioflavin S staining of amyloid fibrils was performed as previously described (21 (link)).