2.9. Terminal Deoxynucleotidyl Transferase-Mediated dUTP Nick 3′-End Labeling (TUNEL)

On day 3 after surgery, mice were transcardially perfused with 0.9% ice-cold saline and 4% paraformaldehyde (Sangon, Shanghai, China) under anesthesia, successively. Then, the corresponding brain samples were collected, fixed in 4% paraformaldehyde for another 1 d, and dehydrated by 25% sucrose for 3 d. The samples were embedded in paraffin and cut into pieces. A TUNEL kit (Beyotime Biotechnology, Haimen, Jiangsu, China) was employed to stain the paraffin pieces according to the manufacturer's protocols.

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