Fig. 2

Fig. 2

Endogenous Nodal signaling requires Vg1.

(A) Mvg1, maternal-zygotic vg1 (MZvg1) and maternal-zygotic oep (MZoep) mutants at 28 hpf. (B) Expression of Nodal target genes gsc, lft1 and ntl at 50% epiboly and sox32 at 90% epiboly in WT, Mvg1 and MZoep embryos. (C) cyc and sqt expression at 4 and 6 hpf in WT and Mvg1 embryos. (D) gsc expression at 50% epiboly in WT and Mvg1 embryos injected with 50 pg of cyc mRNA, 50 pg or 0.2 pg of sqt mRNA. (E) qPCR of lft1 expression at 50% epiboly relative to ef1a in WT and Mvg1 embryos injected with 0.2 pg or 2 pg of sqt mRNA. The mean and standard error of the mean (SEM) was plotted. (F) qPCR of lft1 expression at 50% epiboly relative to ef1a in embryos injected with 20 pg of cyc mRNA in combination with 0.5 pg or 5 pg of vg1 mRNA. The mean and SEM was plotted. (G) Anti-GFP reducing immunoblot of WT and Mvg1 embryos injected with 50 pg of cyc-sfGFP or sqt-sfGFP mRNA. Black arrowhead indicates the position of full-length protein, open arrowhead indicates processed protein. 8 embryos at 50% epiboly were loaded per well. (H) Live imaging of the animal cap of sphere-stage WT and Mvg1 embryos injected with 50 pg of cyc-sfGFP or sqt-sfGFP mRNA. Scale bar, 17 um.