Anti neuronal nuclei neun

Briefly, after PBS perfusion and post-fixation in formalin, brain samples were sectioned into 10-µm-thick slices using a cryostat (CM3050S; Leica Microsystems, Bannockburn, IL, USA). Immunofluorescence staining was performed as previously described (Hasegawa et al., 2011 , Altay et al., 2012 (link)). The sections were incubated overnight at 4°C with the following primary antibodies: anti-Slit2 (1:200), anti-Robo1 (1:200), anti-MPO (1:200) (all from Santa Cruz Biotechnology, Dallas, TX, USA), anti-neuronal nuclei (NeuN) (1:500), anti-glial fibrillary acidic protein (GFAP) (1:100) and anti-CD45 (1:150) (all from Abcam, Cambridge, MA, USA). Sections were then incubated with FITC- and Texas Red-conjugated appropriate secondary antibodies (1:100) (Jackson Immuno Research, West Grove, PA, USA) for 2 hours at room temperature and visualized with a fluorescence microscope (Olympus BX51).