2.9. Western Blot Analysis of Hippocampal Tissue

A portion of hippocampal tissue was placed in a frozen storage tube and immediately stored in liquid nitrogen for detection by western blotting. The total protein and nuclear protein were extracted from myocardial tissue. SDS-PAGE electrophoresis was performed with 50 μg protein. After electrophoresis, the protein was transferred to the NC membrane. Five percent skimmed milk powder was added for blocking at room temperature for 1 h. Next, 5% BSA-diluted antibody was added for incubation at 4°C overnight. The film was washed with TBST 3 times for 15 min each. Next, secondary antibodies in diluent (1 : 1000) were incubated at room temperature for 1 h. Then, the film was washed with TBST 3 times for 15 min each. ECL chemiluminescence color rendering was used, and the film was exposed in a Bio-Rad device (Thermo Fisher Scientific Inc., Waltham, MA, USA) to detect NLRP3, ASC, caspase-1, GSDM-D, IL-1β, IL-6, IL-18, and Aβ protein expression (Cell Signaling Technology, Danvers, MA, USA).