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Fig. 8. Effects of Sprouty2-DN misexpression on Otx2, Gbx2 and Fgf8 expression. In situ hybridization for Otx2 (A-C), Gbx2 (D-F) and Fgf8 (G,H). Misexpression of sprouty2-DN is assessed by immunohistochemistry with the antibody against HA-tag (A,D,G), 24 hours after electroporation (HH 18; A,B,D,E,G,H), 48 hours after electroporation (HH 22; C,F). Sprouty2-DN repressed Otx2 expression, and induced Gbx2 expression in the mesencephalic region. By 48 hours after electroporation, regulation of Otx2 and Gbx2 expression may have occurred, and the expression domain of these genes abuts (C,F). The Fgf8 expression ring is extended rostrally (arrows on G,H). mes, mesencephalon; is, isthmus; met, metencephalon; cont, control side; exp, experimental side. Scale bar: 200 µm.