Immunohistochemistry and immunofluorescent staining was performed using an affinity-purified anti-EGR-1 and anti-pERK1/2 antibodies. The empirical method was identical as previously described [20 (link)]. After dewaxing, rehydration and washing, antigen retrieval was operated in EDTA buffer solution, and the endogenous peroxidase activity was inhibited by incubation with 3% H2O2 for 30 min. Then the sections were incubated with 5% normal fetal bovine serum at room temperature for 30 min. Then primary antibodies (EGR-1, 1:200; pERK1/2, 1:100) were loaded to brain slices (4°C, 12 h). After washing with PBS, peroxidase activity was developed using DAB as the chromogen (Maixin, Fuzhou, China).
The experimental results were photographed under a confocal microscope (×400). Nuclei were counterstained with DAPI, and the stained cells were analyzed and photographed under a confocal microscope(×400).
The experimental results were photographed under a confocal microscope (×400). Nuclei were counterstained with DAPI, and the stained cells were analyzed and photographed under a confocal microscope(×400).
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