2.12 Western blot
After rapid brain extraction, the right cerebral cortex was isolated and
weighed, and the total protein was extracted in a pre-cooled protein
lysate. After quantitative protein electrophoresis, the membranes were
transferred, sealed and blocked for 60 min at 25˚C with QuickBlock
blocking solution (Beyotime Institute of Biotechnology). The primary
antibody solution was refrigerated at 4℃ overnight, and after washing,
secondary antibody was incubated for 2 h at room temperature. Primary
antibodies were diluted as follows; Iba1 (cat. no. BM5765; Boster;
1:1000), IL-1β (cat. no. WL00891; Wanlei; 1:1000), IL-6 (cat. no.
bs-4539R; Bioss; 1:1000), TNF-α (cat. no. WL01581; Wanlei; 1:1000), iNOS
(cat. no. BA0362; Boster; 1:1000), AQP4 (cat. no. A01049; Boster;
1:1000), CD86(cat. no. bs-1035R; Bioss; 1:1000), CD206 (cat. no.
18704-1-AP; ptgcn; 1:1000), Arg1 (cat. no. BM4000; Boster; 1:1000), TLR4
(cat. no. WL00196; Wanlei; 1:1000) and NF-κB (cat. no. WL01980; Wanlei;
1:1000), with β-actin (cat. no. BM0627; Boster; 1:1000) as the internal
reference.