Goat anti iba1

Rabbit kits were anesthetized and intracardially perfused with saline followed by 10% formalin at G29, PND1 and PND5. Brains were post-fixed for 24 h and cryoprotected in 30% sucrose. 30 μm sections were cut with a cryostat and mounted onto poly-L-lysine coated slides (Sigma Aldrich, MO, USA). The sections were blocked with 5% donkey serum, incubated with goat anti-IBA1 (1:250, Abcam, MA, USA), rat anti-CD11b (1:100, Genetex, CA, USA) and mouse anti-CD45 (1:100, Bio-Rad, CA, USA), or goat anti-IBA1 (1:250, Abcam, MA, USA) and rabbit anti-iNOS (1:50, Thermo Fisher Scientific, MA, USA), followed by fluorescent secondary antibodies (1:250, Thermo Fisher Scientific, MA, USA) for 2h. For TSPO and Lectin co-staining, sections were blocked with 5% donkey serum, and incubated with goat anti-TSPO (1:500, GeneTex, CA, USA) overnight at 4°C. The sections were washed, incubated with DyLight 594 lectin (1:250, Vector Laboratory) and fluorescent secondary antibody (1:250, Thermo Fisher Scientific, MA, USA) for 2 h. Sections were washed and incubated in DAPI (1:1000) for 15 min. Immunolabelled sections were cover-slipped with Dako fluorescence mounting medium (VWR, PA, USA). All the images were captured on Zeiss LSM710 confocal microscope, keeping similar settings during image acquisition for different groups.