Skip to main content
. 2016 Apr 14;5:e13758. doi: 10.7554/eLife.13758

Figure 2. Aplnr deficient embryos exhibit a reduction in Nodal signaling.

(A) List and Venn diagram of 23 Nodal target genes found to be down-regulated in a microarray of aplnra/b morphant embryos compared to WT at 50% epiboly (5.25 hpf). (B) Relative luciferase activity regulated by the Activin response element (ARE) in WT, aplnra/b morpholino (MO), oep MO and lefty1/2 MO injected embryos at 30% epiboly (4.7 hpf). Data are represented as means ± SEM. *p<0.05 unpaired two-tailed t-test. (C–E) Phenotypic characterization of WT (D) and aplnra/b morphant embryos (E) when treated with the indicated concentration of the Alk4/5/7 inhibitor SB431542 from the sphere stage (4 hpf) onwards. (F–S) Visualization of the expression of the canonical nodal target genes gsc, flh and sox32 in WT (F,J,N), aplnra/b MOs injected (G,K,O), aplnrains; aplnrbhu4145 double mutant (H,L,P) and aplnrb RNA injected treated with the Aplnr agonist ML233 (I,M,Q) embryos at 8 hpf. Embryos are viewed from the dorsal side. Quantification of the number and spread of sox32 expressing cells (R,S). Data are represented as means ± SEM. *p<0.05, **p<0.01 unpaired two-tailed t-test.

DOI: http://dx.doi.org/10.7554/eLife.13758.006

Figure 2.

Figure 2—figure supplement 1. Loss of aplnr affects Nodal target gene expression.

Figure 2—figure supplement 1.

(A) Gene set enrichment analysis (GSEA) of genes downregulated in aplnra/b morphants when compared against genes upregulated in zebrafish sphere stage (4 hpf) embryos injected with sqt mRNA. (B–G) WISH showing expression of lefty1 at 4 hpf (B, E) and 5.5 hpf (C, F) and lefty2 at 5.5 hpf (D, G) in aplnra/b morphant embryos when compared to WT from an animal pole (top of embryo) view.
DOI: http://dx.doi.org/10.7554/eLife.13758.007
Figure 2—figure supplement 2. Aplnr activation enhances Nodal target gene expression.

Figure 2—figure supplement 2.

(A–I) Expression of the canonical Nodal target genes gsc, flh and sox32 in WT (A–C), aplnrb RNA injected embryos (D–F) and embryos treated with the Aplnr agonist ML233 (G–I) at 8 hpf. Embryos are viewed from the dorsal side with anterior to the top. (J–O) Expression of the canonical Nodal target genes gsc, flh and sox32 in aplnra/b morphant embryos with or without ML233 at 8 hpf. Embryos are viewed from the dorsal side with anterior to the top. (P–U) Expression of the canonical Nodal target genes gsc, flh and sox32 in oep morphant embryos with or without the injection of aplnrb RNA and addition of ML233 at 8 hpf. Embryos are viewed from the dorsal side with anterior to the top.
DOI: http://dx.doi.org/10.7554/eLife.13758.008