FIGURE 6

Effect of Nodal signaling inhibition on ndr1 expression in WT and mutant embryos. WT, Meomesa or Mhwa embryos at the one-cell stage were incubated in Holfreter’s water with 1% DMSO (control) or 50 μM SB431542 (SB) and harvested at indicated stages for detection of ndr1 expression by WISH (A,C,E) or qRT-PCR analysis (B,D,F). Embryos (A,C,E) were positioned in animal-pole view with dorsal to the right if the dorsal side was distinguishable. The ratio of embryos with the representative pattern was indicated in the right bottom. Scale bars, 100 μm. qRT-PCR analysis was performed using 15 embryos per sample. The expression level at 3.7, 4, 4.3, 5 and 6 hpf in WT or Mhwa embryos was normalized to that at 4.7 hpf in WT embryo, while the expression level at different stages in WT or Meomesa embryos was normalized to that at 6 hpf in WT embryos. Error bars indicated S.D. based on three biological replicates (indicated by small circles). Statistically significant levels: ns, nonsignificant; *, p < 0.05; **, p < 0.01; ***, p < 0.001.