Mouse anti syn antibody

Six brain sections at 210–240-μm intervals were extracted from each mouse from the region between −2.6 mm and −4.3 mm to the bregma with reference to Paxinos and Franklin’s the Mouse Brain in Stereotaxic Coordinates [93 ]. To visualize the immunoreactivity of Aβ, NeuN, Iba-1, SYN, GFAP, and pCREB, free-floating sections were incubated overnight at 4 °C with the mouse anti-4G8 antibody (1:2000; BioLegend, San Diego, CA, USA), mouse anti-NeuN antibody (1:100; Merck KGaA, Darmstadt, Germany), goat anti-Iba1 antibody (1:500; Abcam plc, Cambridge, UK), mouse anti-SYN antibody (1:500; Sigma-Aldrich Corporation, St. Louis, MO, USA), rat anti-GFAP (1:200; Thermo Fisher Scientific Inc., Waltham, MA, USA), or mouse anti-pCREB antibody (1:1000; MERCK, Kenilworth, NJ, USA). After washing three times for five minutes in PBS, the sections were incubated with the goat Alexa 488-conjugated anti-mouse IgG (1:200; Thermo Fisher Scientific Inc., Waltham, MA, USA) or donkey Alexa 594-conjugated anti-rabbit IgG (1:200; Thermo Fisher Scientific Inc., Waltham, MA, USA) for 1 h at room temperature. The tissue sections were mounted on ProbeOn™ Plus Microscope Slides (Thermo Fisher Scientific Inc., Waltham, MA, USA) and coverslipped with Fluoroshield™ with DAPI (Sigma-Aldrich Corporation, St. Louis, MO, USA).

Four or seven coronal sections were selected from each mouse from the septal and hippocampal region with reference to Paxinos and Franklin’s The Mouse Brain in Stereotaxic Coordinates [69 ] (Supplementary Figure S3). To examine the immunoreactivity of Aβ, neuronal nuclei (NeuN), and synaptophysin (SYN), free-floating sections were incubated overnight at 4 °C with the mouse anti-4G8 antibody (1:2000; Cat.# 800701, BioLegend, San Diego, CA, USA), mouse anti-NeuN antibody (1:100; Cat.# MAB377, Merk KGaA, Darmstadt, Germany), or mouse anti-SYN antibody (1:500; Cat.# S5768, Sigma-Aldrich), respectively, in blocking solution (0.05% bovine serum albumin, 1.5% normal goat serum and 0.3% Triton X-100 in PBS) [70 (link)]. After washing three times for five minutes in PBS, the sections were incubated with goat Alexa 488-conjugated anti-mouse IgG (1:200; Cat.# A11001, Thermo Fisher Scientific Inc., Waltham, MA, USA) or goat Alexa 594-conjugated anti-mouse IgG (1:200; Cat.# A11005, Thermo Fisher Scientific Inc.) for 1 h at room temperature. The tissue sections were mounted on ProbeOn™ Plus Microscope Slides (Thermo Fisher Scientific Inc.) and coverslipped with Fluoroshield™ with DAPI (Sigma-Aldrich) to counterstain the nuclei.