Anti gad67

Anti-APP antibody (6E10) was purchased from Covance (Cat# SIG-39320-1000). Anti-beta amyloid was purchased from Abcam (Cat#2539). Anti-T. gondii antibody was purchased from US biologicals (Cat# T8075-01). BAG1 was a gift from Dr. Dubey. Normal mouse IgG (SC-2025) and normal rabbit IgG (SC-2027) were used as isotype controls to test the specificity of the 6E10 and BAG1 antibodies, respectively. Anti-phospho-Tau antibody (AT8) was purchased from Thermofisher (Cat# MN1020). Anti-NeuN was purchased from Cell Signaling (Cat# 12943S) or Abcam (ab104224). Anti-VGLUT2 (Cat#71555S), anti-GAPDH (Cat# 2118), and anti-Tau (Cat#4019) antibodies were purchased from Cell Signaling. Anti-NMDAR (Cat# ab17345), anti-VGLUT1 (Cat# ab134283), and anti-GAD67 (Cat# ab26116) antibodies were purchased from Abcam. Thioflavin S was purchased from (Sigma Aldrich) and neuro-tracer from Life Technologies.

Anti-MBP (Aves Labs), anti-GFAP (DAKO), anti-F4/80 (clone Cl:A3-1; AbD Serotec), anti-PSD-95 (Abcam), anti-vGlut2 (Abcam), anti-GAD67 (Abcam), and Alexa488- and/or Alexa594-conjugated secondary antibodies (Molecular Probes) were used for immunohistochemistry. Anti-GluN2B (Synaptic Systems) antibodies were used for Western blot analysis and immunohistochemistry. Anti-GAPDH (EMD Millipore) and horseradish peroxidase (HRP)-labeled secondary antibodies (Vector Laboratories) were used for Western blot analysis.

According to the instructions, the total protein was extracted from the globus pallidus with a total protein extraction kit (KGP250; KeyGen Biotech, Nanjing, China), and a bicinchoninic assay (BCA) protein detection kit (KGPBCA; KeyGen Biotech, Nanjing, China) was used to quantify the protein concentrations. Target proteins were isolated using sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and subsequently transferred to a polyvinylidene difluoride (PVDF) membrane. After cleaning with Tris-buffered saline with a Tween 20 (TBST) solution, the PVDF membranes were sealed in a blocking solution for 15 min. They were incubated overnight at 4 °C with anti-GAD67 (1:1000; Abcam, Cambridge, UK), anti-caspase-3 (1:1000; Abcam, Cambridge, UK) and anti-β-actin (1:2000; ABclonal, Wuhan, China) antibodies. The next day, the PVDF membranes were washed with TBST and incubated with the respective secondary antibodies for 1 h. Subsequently, each PVDF membrane was treated with an enhanced chemiluminescence (ECL) solution and exposed using the Bio-Rad ChemiDoc Touch System (Bio-Rad Laboratories, Hercules, CA, USA). Image analysis was performed using ImageJ software (Version 1.53; NIH, Bethesda, MD, USA).