Figure 2

scfd1 deficiency is responsible for the hah^vcc43 mutant phenotype. (A) Brightfield photographs show similar phenotypes in hah^vcc43−/− mutants (middle left), scfd1 morphants (middle right), and scfd1^vcc44−/− mutants (right) in comparison to wildtype (WT; (+/+; left) at 3 days post fertilization (dpf). These include cardiac effusion and poorly ballooned chambers (red arrow), as well as craniofacial defects such as an absence of lower jaw (blue arrowheads) in comparison to a clearly defined jawline in +/+ (blue outline) and small eyes (green asterisk in dotted circle). (B) Both hah^vcc43 and scfd1^vcc44 mutants harbor a PTC in exon 10 of scfd1. (C) Bar graph showing the proportion of hah embryos exhibiting normal (black bars), mild (dark gray bars), or severe (light gray bars) mutant phenotypes at 4 dpf after injection with WT scfd1 mRNA. Note reduction in proportion of affected embryos with increasing concentrations of scfd1 mRNA. See Methods for definition of phenotypes. Numbers of embryos in each group indicated at base of bar. Chi-square p-values stated. (D,E) Scatter plots showing reduced scfd1 transcript expression as determined by qRT-PCR (D; 30 embryos/sample, n = 5 samples) and protein expression as determined using Western blot (E; 30 embryos/sample, n = 6 samples) in both heterozygous (+/−) and homozygous (−/−) hah^vcc43 and scfd1^vcc44 mutants at 3 dpf. Ordinary one-way ANOVA p-values stated. Significance if p < 0.05. α-tubulin loading control used for normalization for Western blot quantification. Full blots are shown in Supplementary Figure S2. Created with BioRender.com.