*4.7. Histological Analysis*

Before initiation of brain tissue immunostaining, blocking of endogenous peroxidase activity is important. The previously described activity was blocked by 1.2% hydrogen peroxide incubation for 20 min at room temperature. After incubation and washing in 0.01% phosphate buffered saline, sections were incubated with several antibodies to analyze our data. In this study, the antibodies used were as follows: 4HNE (diluted 1:500, Alpha Diagnostic Intl. Inc., San Antonio, TX, USA), GFAP (diluted 1:1000, Abcam, Cambridge, UK), Iba-1 (diluted 1:500, Abcam), TRPM2 (diluted 1:400, Abcam), and NeuN (diluted 1:500, Billerica, Millipore Co., Burlington, MA, USA).
