Rabbit polyclonal anti c fos

To assess overall neuronal activity of the DRN immunohistochemical expression of c-Fos was evaluated. The DRN sections were incubated overnight with a primary anti-c-Fos antibody (rabbit polyclonal anti-c-Fos; Abcam; 1:1000). This was followed by incubation for one hour with a secondary antibody (donkey anti-rabbit alexa 594, Jackson immunoresearch Laboratory; 1:200). Eight slices were selected from bregma − 4.16 to − 4.96 and photographed with an Olympus BX51 fluorescence microscope (Olympus, Germany) connected to an Olympus Camera DP72 (Olympus, Germany). All clear c-Fos expressing neurons were counted (FiJi v2.0.0, National Institutes of Health; Maryland).

High-fat diet (D12451, 45% kcal fat) and normal diet (D12450B, 10% kcal fat) were purchased from Research Diets, Inc. (New Brunswick, NJ, USA). Rabbit polyclonal antipost synaptic density protein 95 (PSD95) and rabbit polyclonal anti-c-Fos were purchased from Abcam (Cambridge, UK). Biotinylated goat anti-mouse antibody, biotinylated goat anti-rabbit antibody, and avidin-biotin complex (ABC) were purchased from Vector Labs, Inc. (Burlingame, CA, USA). Mouse monoclonal anti-synaptophysin (SYN), paraformaldehyde (PFA), 3,3-diaminobenzidine (DAB), sodium chloride, sucrose, ethanol, and phosphate buffered saline (PBS) were purchased from Sigma-Aldrich (St. Louis, MO, USA).