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