Generation and morphology of epm2a−/− zebrafish mutants. (A) Coding sequence alignment of wt and mutant epm2a shows the generation of epm2a-null mutant zebrafish by insertion of 5 bp into exon 2 of the epm2a gene. (B) Protein alignment of wt and mutant epm2a. The red rectangle shows the premature stop codon at residue 74 (p.Thr54Asnfs*74) caused by the 5-bp insertion mutation in exon 2. (C) qRT–PCR analysis revealed a decrease in the level of epm2a mRNA expression, normalized to the expression of β-actin mRNA. Three independent RNA samples (each obtained from about 30–40 larvae) from epm2a−/− larvae at 120 hpf and from controls were analyzed. The values are expressed as mean ± standard deviation (SD). **, p ≤ 0.01, calculated by Student’s t-test. (D) Lateral view photographs of representative control and epm2a−/− specimens. At 4 dpf we observed a significant reduction in body length in epm2a−/− larvae (n = 83) compared to WT controls (n = 83), **** p ≤ 0.0001, calculated by Mann-Whitney test.
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