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Figure 1.
Rapgef5 Depletion Disrupts LR Development
(A) Percentage of Rapgef5 depleted embryos with abnormal cardiac looping (A or L loops).
(B) Pitx2c is expressed in the left lateral mesoderm of stage 28 control Xenopus embryos (red arrow, lateral view anterior to the left), but is abnormally, typically bilaterally, expressed following MO- or CRISPR-mediated depletion of Rapgef5. Co-injection of human RAPGEF5 mRNA can rescue pitx2c in Rapgef5 morphants.
(C) coco expression in the LRO of control and Rapgef5-depleted embryos at stages 16 and 19. Ventral view with anterior to the top. Graphs depict the percentage of embryos displaying abnormal coco expression. Note the reduced expression in rapgef5 morphants at both stages.
(D) xnr1 and gdf3 expression is reduced in the LRO of rapgef5 morphants at stage 16, prior to the onset of cilia-driven flow. Ventral views with anterior to the top.
∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.005. See also Figures S1 and S2. |
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Figure 5.
NLS β-Catenin Rescues foxj1 Transcription and LR Patterning Defects in Rapgef5 Morphants
Depletion of Rapgef5 reduces the expression of the Wnt responsive gene foxj1 and alters LR patterning (pitx2c). Red arrow indicates an example of normal pitx2c expression on the left side of the embryo. Co-injection of 50 pg of NLS-β-catenin mRNA significantly rescues (A) expression of foxj1 at stage 10 and (B) pitx2c expression in Rapgef5 depleted embryos. ∗p < 0.05, ∗∗p < 0.01. |
