The frequency of Immunohistochemistry (IHC) staining was performed as described29 (link). To identify CD8+ cells in brain, anti-CD8α (1:400, Clone D4W2Z, Cell Signaling Technology) was used. Images were obtained with a Zeiss Axio Imager Z1. The number of cells showing CD8 positivity in each individual high-power field (HPF) (× 200) was quantified across five fields or more of nonsequential cryosections 9 mm in thickness.
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