Quantifying α7nAChR Expression in Brain Tissues

The expression of α7nAChR was evaluated after a 4-week estrogen pre-treatment. According to Guo et al. (2013) (link), brain tissue proteins were extracted. After subjecting equivalent protein amounts (50 μg) to SDS-PAGE, they were then transferred to a polyvinylidene fluoride membrane. After being blocked for 2 h with 5% BSA at room temperature, the membranes were incubated with an anti-α7-nAChR antibody (1:1000) or an anti-GAPDH antibody (1:20,000) before incubation with IRDye 800CW-conjugated secondary antibody (Rockland Immunochemicals, Inc.). The Odyssey Infrared Imaging System (Li-Cor Biosciences, Lincoln, NE, United States) was utilized for capturing the images, and ImageJ software (NIH) was used for analyzing them.