Immunofluorescence Staining of Brain Sections

Immunofluorescence staining was performed according to our previously published Protocols (Sun et al., 2014 (link); Ye et al., 2018 (link)). Briefly, brain sections (6 μm) were first incubated with primary antibodies against p65 (1:200, 1:1000, cat. No. D14E12, CST), iba-1 (1:200, RRID: AB_2224402) overnight at 4 °C. The next day, brain sections were incubated with corresponding secondary antibodies Alexa Fluor 488 and/or Alexa Fluor 594 (Jackson ImmunoResearch Incorporation, West Grove, PA, United States). Fluorescence was examined under a ZEISS HB050 inverted microscope system. The fluorescently stained cells were analyzed by Image J software.