The establishment of the zebrafish elmo1 mutant. (A) Schematic representation of the elmo1 genomic locus (NC_007130.7). The extended region on exon 18 represents the sequence targeted by the TALEN system. Green: left TALEN arm binding site. Blue: right TALEN arm binding site. Red: spacer site. elmo1+/+ corresponds to the wild-type allele while elmo1−/− represents the loss of function allele in this study. Dashes represent the 13 base pair deletion. (B) Schematic view of the wild-type Elmo1 protein (Elmo1 wt) and the mutated Elmo1 protein (Elmo1 mut). The 726 amino acid (aa) Elmo1 wt contains five conserved domains (NP_998,256), while the Elmo1 mut resulting in a truncated protein end up at 619 aa. RBD: RhoG-binding domain; EID: ELMO inhibitory domain; ELMO1: ELMO1 domain; PH: pleckstrin homology; EAD: ELMO auto-regulatory domain. (C) The qRT-PCR result showed the relative expression level of the elmo1 gene in the offsprings of heterozygous intercross at 3 dpf. Three independent experiments were performed. One-way ANOVA, *p < 0.05, ***p < 0.005. (D) elmo1 expression pattern detected by WISH in the offsprings of heterozygous intercross at 3 dpf. Scale bar: 200 μm.
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