Fig. 3
Vg1 processing requires Nodal.
(A) superfolderGFP (sfGFP) was inserted into vg1 downstream of the predicted basic cleavage site. (B) Mvg1 embryo injected with 50 pg of vg1-sfGFP mRNA, shown at 28 hpf. (C) Anti-GFP reducing immunoblot of WT and Mvg1 embryos injected with 50 pg of vg1-sfGFP mRNA. Black arrowhead indicates full-length Vg1-sfGFP; open arrowhead indicates the predicted size of cleaved Vg1-sfGFP. 8 embryos at 50% epiboly were loaded per well. (D) Mvg1 embryo injected with 50 pg of non-cleavable vg1 mRNA (vg1-NC, RSRRKR->SQNTSN), shown at 28 hpf. Embryos were injected with up to 200 pg of vg1-NC mRNA with no rescue. (E) Anti-GFP reducing immunoblot of Mvg1 embryos injected with 10 pg of vg1-sfGFP or vg1-NC-sfGFP mRNA and 10 pg of cyc or sqt mRNA. Black arrowhead indicates full-length Vg1-sfGFP, open arrowhead indicates cleaved Vg1-sfGFP. Molecular weights in kDa. 8 embryos at 50% epiboly were loaded per well. See also Figure 3—figure supplement 1.