(A) Expression of α-synuclein was examined at the indicated times after cell dissemination. Immunoreactivity of α-synuclein increased during cell maturation. By 30 h after cell dissemination, some NeuN-positive cells expressed α-synuclein (asterisks), but other NeuN-positive cells did not. (B) Expression of GAD at 30 h was not sufficient to distinguish inhibitory neurons from other types of neurons. Three independent cultures were performed. Scale bars: 10 µm.