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Stereological brain sampling of the areas of interest was performed and brain were serially cut on a cryostat (Leica Biosystems, Germany, cat# CM3050S), mounted on Superfrost Plus slides (Thermo Fisher Scientific, Reinach, Switzerland), and cryoprotected in 50% of PB, 30% of ethylene glycol, 20% of glycerol, and subsequently stored at 20 C