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Fig. 5
Mutant amer2 and amer3 alleles are not associated with embryonic anomalies. A, Relative mRNA levels of wtx, amer2 and amer3 in female ovaries and in zebrafish embryos of different developmental stages (1 hpf ‐ 48 hpf) of wild type and homozygous wtx mutants ( wtx del1 , wtx del3 ), analyzed by qRT‐PCR. Gene expression was normalized to ef1a. Wild type was set as 1; the ratio between mutant/wild type is shown. N = 3 for wild type and wtx del3 ovaries, as well as for pooled embryos (expression data of wtx del1 and wtx del3 were combined). The error bars indicate SEM. B, Structure and partial sequence of the amer2 gene locus on zebrafish chromosome 24 and the amer3 gene locus on zebrafish chromosome 2. The coding sequences are indicated as black boxes, grey boxes depict untranslated sequences. Binding sites of the TALENs are indicated by arrowheads. Below the schemes sequences of selected amer2 and amer3mutations in F1 fish are shown. Deletions are indicated by dashes. Red letters indicate nucleotide insertions. Underlined sequences correspond to the left and the right arm of the TALENs. In brackets the total number of deleted or inserted nucleotides in mutant alleles are indicated. C and D, Representative brightfield images of 7 dpf homozygous mutant zebrafish larvae resulting from an in‐cross of heterozygous fish with TALEN induced B, amer2 del2 and C, amer3 del1 mutation. E, Relative mRNA expression of wtx, amer2 and axin2 in uninjured fin tissue at 3 dpa measured by qRT‐PCR. Gene expression was normalized to uninjured fin (0 dpa) and to the reference gene ef1a. Tissues of three animals were pooled. Error bars represent SEM