Immunofluorescence Staining of Neural Markers

Immunofluorescence staining was performed as we described in detail before [17 (link)]. In brief, the sections were first fixed by 0.1% Triton X-100 and then were blocked by 5% BSA for 2 h. After three washes with PBS, brain tissues or cultured neurons were incubated with primary antibodies against Iba-1 (1: 50, Santa Cruz Biotechnology or 1 : 100, Abcam), myeloperoxidase (MPO, 1 : 50, Santa Cruz Biotechnology), 8-hydroxydeoxyguanosine (8-OHdG) (1 : 100, Abcam), Nrf2 (1 : 100, Abcam), caspase-1 p20 (1 : 50, Santa Cruz Biotechnology), and NeuN (1 : 200, EMD Millipore). Sections were then incubated with corresponding secondary antibodies (Alexa Fluor 488 and Alexa Fluor 594) overnight at 4°C. Fluorescence microscopy imaging was examined under a ZEISS HB050 inverted microscope system. The fluorescently stained cells were recorded using Image J program.