hace1?morphant embryos display specific cardiac phenotypes that can be rescued by human HACE1 mRNA while the vasculature remains intact. A: The looping defect that results from hace1 knockdown results in either misalignment of the atrium and the ventricle (the ventricle is located on the left side [white arrow], ?inverted?) in ii or a tubular heart (?straight') in iii. Scale bar?=?250?µm. B: Diagrammatic representation of each phenotype is included adjacent to a fluorescent image of a Tg(myl7::eGFP) hace1?morphant embryo with the associated phenotype. C: Bar graphs showing the percent of each of the representative anatomic phenotypes shown in panels A and B (colors correspond to the respective phenotypes; n?=?90?130 embryos for each group in 3 replicates; lateral views at 48 hpf; **P?0.0001). D: Bar graphs showing the heart rate in hace1 and control morpholino?injected embryos under the conditions indicated (n?=?30?40 embryos for each group; * denotes a significant difference between the two groups of hace1 morphants, which also differ significantly with all other groups included in this panel at *P?0.05). Movies capturing the differences in heart rates can be found in Supp. Movie S1. E: Fluorescence imaging of the 48?hpf Tg(fli1a::eGFP) embryos injected with either control or hace1 morpholino demonstrates an intact vascular structure (lateral views) with a nonsignificant (ns) difference between the percentage of embryos with normal vasculature shown in F (n?=?3, P?>?0.05). Scale bar?=?250?µm.
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