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Still heart mutants lack hand2-migrating cells and hand2 expression in their hearts. At 24 hpf, hand2 expression can be observed in the bilateral cell populations of the anterior lateral plate mesoderm in both wild-type (A) and still heart mutants (B). By 36 hpf, hand2 expression has expanded toward the midline (dotted white line) of wild-type embryos (C), while there is a notable void of hand2 expression proximal to the midline in still heart mutants (D). At 48 hpf, wild-type embryos display hand2 expression in their pharyngeal arches (PAs) and heart (H) (E). Still heart mutants demonstrate hand2 expression in their pharyngeal arches but lack hand2 expression in their hearts (F). Comparison of relative hand2 expression in wild-type and still heart mutants revealed no significant difference in hand2 expression at 24 hpf (G). However, by 48 hpf, still heart mutants demonstrate a significant reduction in the relative fold change in hand2 expression (H). *** = p ≤ 0.001.
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