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Ebf2, Ebf3, and Stmn4 mRNA are expressed in DA neurons in the PT area of the diencephalon. Larvae are subject to double-labeled in situ hybridization followed by coronal sections. A: Double-labeled in situ hybridization reveals that Ebf2 (red) and TH (green) are only colocalized in a subgroup of DA neurons at 24 hours postfertilization (hpf). Most Ebf2 signals surround TH signals. Lower panels are the enlarged views of the stippled boxes in the upper panels. B: Double-labeled in situ hybridization of Ebf3 and TH. They also show co-expression in a subgroup of DA neurons at 24 hpf. C: Double-labeled in situ hybridization of Stmn4 and TH. Colocalization is observed at 24, 30, and 48 hpf. Arrows in A, B, C indicate the cells with colocalized hybridization signals. DA, dopaminergic; TH, tyrosine hydroxylase; PT, posterior tuberculum.
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