Anti cd16 32

Western blots were performed using a previously reported method [19] . The cerebral cortex and hippocampus on the injured side were separated and stored in liquid nitrogen until use. Total proteins were extracted, and then the protein concentration was measured using a BCA protein assay kit (Solarbio Science, Beijing, China). Protein samples were separated by SDS-PAGE and subsequently transferred onto PVDF membranes (Millipore, Bedford, MA). The blots were blocked with 5% non-fat milk for 2 h at room temperature, then immersed overnight at 4 ℃ in a solution containing the primary antibodies: anti-TLR4 (1:500, GeneTex, USA), anti-APP (1:500, CST, USA), anti-p-Tau(1:500, CST, USA), anti-MyD88 (1:1000, CST, USA), anti-NF-κB p65 (1:1000, CST, USA), anti-CD16/32 (1:1000, Abcam, MA, USA), and anti-CD206(1:1000, Abcam, MA, USA). Afterwards, membranes were washed with TBST and incubated for 1 h at room temperature with HRP goat anti-mouse and HRP goat anti-rabbit secondary antibodies (all diluted 1:5000, ZhongShan, Beijing, China). The blots were treated with enhanced chemiluminescence (ECL) reagents and visualized with a ChemiDoc TM XRS + Image System (Bio-Rad, CA, USA). The intensity of each band was analysed using ImageJ software.