Effects in embryos co-injected with MOs against both Chl and Chd demonstrate overlapping roles for these genes in dorsoventral patterning. A comparison is shown at 24 hpf of embryos treated with 0.5013 mM StCon-MO (A), 0.1 mM Chl 5′1-MO plus 0.0013 mM StCon-MO (B), 0.5 mM Chl SP-MO plus 0.0013 mM StCon-MO (C), or 0.026 mM Chd-MO alone (D) or with combinations of 0.1 mM Chl 5′1-MO plus 0.026 mM Chd-MO (E) or 0.5 mM Chl SP-MO plus 0.026 mM Chd-MO (F). Single injections of 5′1-MO (B) and SP-MO (C) had added StCon-MO to match total amounts of MOs used in the co-injection experiments (E and F). (G) In situ hybridization shows altered expression patterns at 30% epiboly for the dorsal marker Chd (top panel) and at 70% epiboly for the ventral marker Gata2 (bottom panel) in response to treatment of embryos with the Chd-MO (CHD-MO) and Chl 5′1′MO (Chl-MO), in comparison to embryos treated with StCon-MO (Cont-MO). Concentrations of MOs used were the same as those used in panel A. Note the increased reduction of the Chd expression domain and enlargement of the Gata2 expression domain in response to co-injection with the Chd and Chl-MOs.
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