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Later the ethanol fixed cells were washed with PBS and incubated for 15 min in staining buffer which contains triton-X-100, PI, and RNAse enzyme) and then cells were vortexed and analyzed by flow cytometry (BD C6 Accuri flow cytometry, USA)

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Additionally, these samples exhibited brain matrix vacuolization, increased reactive astrocytic and microglial infiltration (arrowhead), and severely congested blood vessels (star)