Rat anti mouse cd86

Spinal cord sections mounted on slides for immunofluorescence were rinsed in PBS for 5 min to remove residual OCT. After washing, samples were blocked in serum of the host of the secondary antibody (5% serum and 0.3% BSA in PBS with 0.2% Triton-X 100) and then incubated overnight with rabbit anti-mouse Iba1 (1:500, Wako), mouse anti-mouse iNOS (1:500, BD Biosciences), mouse anti-mouse Arg-1 (1:500, BD Biosciences), rat anti-mouse CD86 (1:500, BD Biosciences), goat anti-mouse CD206 (1:100, R&D Systems), rabbit anti-mouse GFAP (1:1000, Dako) rabbit anti-mouse NG2 (1:500, a generous gift from the Levine lab), mouse anti-mouse CC1 (1:100, EMD Millipore), rat anti-mouse MBP (1:50, Bio-Rad), or rabbit anti-mouse neurofilament-L (1:100, EMD Millipore) in 0.3% BSA in PBS with 0.2% Triton-X 100. After washing with PBS, sections were incubated with fluorescence-conjugated FITC or Cy3 goat anti-rabbit, goat anti-mouse, goat anti-rat, or donkey anti-goat antibody for 1 h at room temperature, washed three times with PBS, and mounted using Fluoromount-G with DAPI (Southern Biotech, USA). The sections were imaged using a Nikon Eclipse E600 microscope and Zeiss LSM 510 confocal microscope.