FIGURE 3

Induction of nodal genes and mesendodermal markers in Meomesa;Mhwa mutants by ectopic hwa or/and eomesa. One-cell stage mutant embryos were injected with corresponding mRNA or/and MO and observed for morphology at 6 hpf (A) or harvested at indicated stages for detection of selected genes by WISH (B,D) or by RT-PCR analysis (C,E). Embryos were positioned laterally (A) or in animal-pole view (B,D) with dorsal to the right if the dorsal side was perceptible. The ratio of embryos with the representative pattern was indicated in the right bottom. Scale bars: 100 μm. Injection doses of mRNA or MO: hwa and myc-eomesa, 50 pg/embryo; cMO (as control MO) and β-cat2-MO, 20 ng/embryo. qRT-PCR analysis was performed using 15 embryos per sample, and the expression level was normalized to that of eif4g2a in WT embryos at the same stage. Error bars indicated S.D. based on three biological replicates (indicated by small circles). Color keys for embryo types and treatments were shown in (A,B,D). Statistically significant level: ns, nonsignificant; *, p < 0.05; **, p < 0.01; ***, p < 0.001.