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Fig. 4 Knockout of Zebrafish arl4aa Perturbed Definitive Hematopoiesis in F2 Mutants (A) Strategy of raising F2 mutant embryos. (B) DNA sequencing and genotyping of F2 mutant embryos containing 2-bp deletion and 6-bp substitutions. The amino acids are labeled under the DNA sequence. (C) Expression of c-myb in F2 embryos (upper, WT; middle, heterozygous; lower, homozygous) at 36 (left panel) and 48 hpf (middle panel) and of rag1 at 96 hpf (right panel). (D and E) Intensity of c-myb expression in CHT at 48 hpf (D) and rag1 expression in thymus at 96 hpf (E) as measured by ImageJ software. Data are shown by means ± SEM; n and p values are indicated in (D) and (E). Scale bars, 250 μm (in WISH). The numbers in each WISH showed the numbers of embryos with typical gene expression as shown over the total number of embryos injected in each group.