Phylogenetic and expression analysis of Eif2b orthologs in zebrafish during development. (A) A horizontal cladogram of eif2b1-5 gene sequences shows that zebrafish have a single ortholog of each human EIF2B gene, and a relative conservation of amino acid sequence between orthologs. Scale bar equals an evolutionary distance of 0.1 amino acid changes per position in the sequence (Phylodendron). (B) Comparison of zebrafish and human EIF2B protein sequences shows conserved domain architectures. (C–L) Whole-mount in situ expression analysis in zebrafish embryos and larvae, brightfield microscopy, rostral left, dorsal top. (C–E) eif2b subunit genes at 24 hpf shows predominantly brain and eye expression, with lower levels of expression throughout the embryo. (H–L) eif2b subunit genes at 72 hpf shows higher expression throughout the animal and in the brain compared to 24 hpf. (M–Q) qRT-PCR expression of eif2b subunit genes from 24 hpf through 7 dpf, normalized to 24 hpf expression demonstrates variable expression changes across development. Error bars, standard error of the mean.
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