Standard immunohistochemistry procedures were described previously 53 (link), 59 (link). The samples harvested at 5 days were labeled with the following primary antibodies: anti-S-100 (labeling SCs, 1:1000) and anti-LC3 (labeling autophagy, 1:1000), anti-MPZ (labeling myelin, 1:1000) and anti-GFAP (labeling dedifferentiated SCs, 1:500), anti-GFAP and anti-p75NTR (NGF receptor, 1:1000). The samples collected at 14 days were double-stained with anti-NF-200 (labeling axon, 1:100,000) and anti-MBP (myelin marker, 1:1000). The sections were sequentially incubated with FITC-conjugated anti-rabbit IgG (Abcam, ab150073) or TRITC-conjugated anti-mouse IgG (Abcam, ab7065). The statistical indicators of the immunofluorescence staining were measured in three randomly selected fields from one section, and a total of six sections from three animals of each group was used for capturing images with a Nikon confocal laser microscope (Nikon, A1 PLUS, Tokyo, Japan) or a Nikon Eclipse 80i fluorescence microscope. For different antibody immunoreactivity positive areas (%), the calculation formula used was IOD / selected region area*100%. The IOD and selected region area were measured using the Image pro-plus software.
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