Hippocampal α7nAChR Protein Expression

The extraction of hippocampal proteins was performed with the assistance of ice-cold RIPA lysis buffer (Beyotime, Shanghai, China), which includes a protease inhibitor (Beyotime, Shanghai, China) and a phosphatase inhibitor (Boster, Wuhan, China). As previously reported, we employed the Western blot technique to assess the expression of the α7nAChR subunit [22 (link)]. The membranes were incubated with primary antibodies against rabbit α7nAChR (Abcam, ab216485, 1 : 1000) and against mouse ß-actin (Boster, Wuhan, China, BM0627, 1 : 500) then incubated with homologous secondary antibodies. Enhanced chemiluminescence (Bio-Rad) was employed with the goal of visualizing the protein bands on the membranes. Subsequently, the Image Lab program (Bio-Rad, Richmond, CA, USA) was used to scan the protein bands in each group and determine their relative density in each group. With the aid of the NIH Image J program (Bethesda, MD, USA), we successfully determined the intensity of the bands.