cells were mounted in PermaFluor Aqueous Mounting Medium (Thermo Fisher Scientific, Waltham, MA, USA). Fluorescent signals were detected using a confocal laser-scanning microscope FV1000 (Olympus, Tokyo, Japan). We counted cells positive for both βIII-tubulin and DAPI signals as neurons. These signals can differentiate each cell as βIII stains neuronal cell bodies and neurites, and DAPI stains nuclei. Astrocytes were defined as both GFAP- and DAPI-positive.
