Af7569

At the end of the Morris water maze test (n =6), total protein was extracted from hippocampal tissues under anesthesia with sevoflurane. SDS-PAGE (8%) for NF-κB and SDS-PAGE (12%) for IL-6 and IL-1β were used to perform electrophoresis. Then, the protein was shifted onto a PVDF membrane. After blocking, the PVDF membranes were processed by rabbit anti-rat phospho-NF-κB (1:1000 dilution, AF2006; Affinity Bioscience, OH, USA), rabbit anti-rat polyclonal NF-κB (1:1000 dilution, AF7569; Beyotime, China), mouse anti-rat Monoclonal IL-6 (1:1000 dilution, AF0201; Beyotime, China), and rabbit anti-rat polyclonal IL-1β (1:500 dilution, K101295P; Solarbio, China) at 4°C overnight. Next day, the PVDF membranes were processed by corresponding secondary antibodies (goat anti-rabbit antibody, 1:1000 dilution, A0516; Beyotime; goat anti-mouse antibody, 1:1000 dilution, A0521; Beyotime) at 25°C for 1 h. In addition, the internal reference was performed by a polyclonal rabbit anti-rat GAPDH (1:1000 dilution, K200057M; Solarbio, Beijing, China). Following rising with TBS-T and incubation with ECL (P0018AM; Beyotime), we quantified protein bands on the PVDF membranes with Image-Pro Plus 6.0 software.