Ab12274

Immunohistochemistry was performed on paraffin-embedded ascending aorta sections. Slices were incubated with anti-TGF-β1 (ab92486, Abcam, USA), anti-MMP9 (bs-4593R, Bioss, China), anti-GAP43 (ab12274, Abcam, USA), and anti-TH (ab112, Abcam, USA) overnight at 4°C. The slides were reacted with peroxidase conjugated goat anti-rabbit IgG (ZDR-5306, Zhongshan, China) at 37°C for 20 min [15 (link), 27 (link)].

Western blot analysis was carried out in brain tissue homogenates as reported previously [36 (link)]. Briefly, brain samples were homogenized in lysis buffer. Total protein content was determined using a Pierce™ BCA Protein Assay Kit (Thermo Scientific). Band analysis was performed using ChemiDoc XRS system (BioRad, Munich, Germany). The following antibodies were used: full-length AβPP (fl-AβPP) and C-terminal fragments (CTFs) were detected using AβPP C-Terminal Fragment (C1/6.1) antibody (SIG-39152; Covance, Princeton, NJ, USA). Detection of respiratory system complexes was carried out by MitoProfile^® Total OXPHOS Rodent WB Antibody Cocktail (ab110413; abcam, Cambridge, UK). As primary antibody for synaptic markers, Anti-BDNF (ab72439; abcam), Anti-GAP43 (ab12274; abcam), and Anti-synaptophysin (ab32127; abcam) were used. To detect protein expression of PGC-1α in the brain, Anti-PGC-1α antibody (ab106814; abcam) was used. Anti-tubulin antibody (ab6160; abcam) and Anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody (MAB374; Merck Millipore, Darmstadt, Germany) were used to verify equal protein loading.