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Fig. S6
Analysis of zygotic and maternal zygotic null gdf3 embryos, Related to Figure 5.
(A-C) Brightfield images of 38 hours post-fertilization (hpf) control (CTRL; n=20), zygotic null gdf3 (Zgdf3; n=7), and maternal zygotic null gdf3 (MZgdf3; n=25) embryos processed by in situ hybridization with a tie1 riboprobe. The numbers identify each PAA cluster in (A) and (B). The asterisk in (C) highlights endothelial cells of unknown identity. Lateral views, anterior left. (D-I) Confocal z-stacks of 72 hpf CTRL (n=14), Zgdf3 (n=11), and MZgdf3 (n=32) embryos double immunostained with antibodies recognizing striated muscle (MF20, red) and OFT smooth muscle (Eln2, green). Arrows in (D,E) highlight the Eln2+ OFT smooth muscle that is missing in MZgdf3 mutants. Nuclei were counterstained with DAPI in (F,I). Merged double (D,E,G,H) and triple channel (F,I) images are shown. MZgdf3 animals are devoid of head and trunk mesoderm and therefore lack striated and smooth muscle. Ventral views, anterior up in (D-F). Lateral views, anterior left in (G-I). (J-L) Brightfield images of 6 somite stage (ss) CTRL (n=21), Zgdf3 (n=6), and MZgdf3 (n=26) embryos processed by in situ hybridization with an nkx2.5 riboprobe. Dorsal views, anterior up. In all cases, little to no variation was observed between animals in each experimental group. Abbr: LDA, lateral dorsal aorta; PA1-derived head muscles: ima, intermandibularis anterior; imp, intermandibularis posterior; am, abductor mandibulae; lap, levator arcus palatini; do, dilator operculi; PA2- derived head muscles: ih, interhyoidus; hh, hyohyoidus; ah, adductor hyoideus; ao, adductor operculi; lo, levator operculi; OFT, outflow tract. Scale bars=25 μm.