For TUNEL assay, cryopreserved sections were fixed in 4% PFA for 20 min at room temperature and incubated with sodium citrate solution containing 0.3% Triton X-100 for 30 min. Subsequently, TUNEL reaction mixture (Roche, 11684795910) was added and incubated with brain sections for 1 h at 37 °C in the dark. After staining, sections were coverslipped with a mounting medium and observed under a microscope.

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