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Fig. 7. iro1 and iro7 act together as repressors in formation of the MHB domain, while ectopic iro1 or iro7 induces the ectopic expression of MHB genes. (A-E) The effects of the iro1 (MO1) and iro7 (MO7) morpholinos on expression of MHB markers eng2 (A), pax2.1 (B), fgf8 (C), wnt1 (D) at 24 hpf and pax2.1 (E) at tailbud stage. Red arrows show reduction of MHB markers. (F) Expression of pax2.1 in hdl mutants (left) is expanded to the anterior but its expression is reduced in MO1 and MO7 co-injected hdl mutants (right). (G) Expression of pax2.1 is recovered (red arrowhead) by {Delta}Niro1 and {Delta}Niro7 mRNA co-injection in double morpholino-injected embryos. (H-J) Embryos injected with iro1 mRNA or En-iro7HD mRNA showed ectopic expression of pax2.1 and fgf8, but not wnt1 (F,H), while iro7 mRNA induces fgf8 and reduces wnt1 (G). The distribution of injected mRNA is marked by red staining. (K-M) VP16-iro7HD mRNA injected embryos show loss of pax2.1 expression (K) at tailbud stage and the isthmus at the MHB region (L, arrowhead) and loss of fgf8 expression (M) at 24 hpf. Anterior is towards the left (A-M). Embryos are viewed from the left side (A-D,M) or dorsal (E-L).