Nissl staining was performed to evaluate neurons as described previously (Zhang et al., 2012 (link); Chen et al., 2013 (link)). After washing with PBS, the sections were incubated with Nissl Staining Solution (Beyotime Institute of Biotechnology, Nanjing, China) for 20 min at room temperature. Sections were then dehydrated with ethanol and covered with Permount.
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