Fig. 1

Generation of mutant hspb7 zebrafish lines. (A) Structure and partial sequence of the hspb7 gene, showing TALEN targeting proximal to the start codon and resultant mutant alleles WC3 and WC4. (B) qPCR data showing that hspb7^WC3 and hspb7^WC4 transcripts are expressed at levels comparable to wildtype hspb7 in 5 dpf zebrafish embryos. (C) Western blot of protein lysates from adult zebrafish hearts showing loss of Hspb7 protein in animals homozygous for either of the hspb7 mutant alleles. (D) RFLP analysis showing loss of the HpyCH4V cut site in PCR product from hspb7^WC3/WC3 embryos. Quantification of embryonic genotypes demonstrates that hspb7^WC3/WC3 alleles are transmitted at Mendelian ratios. (E) Representative brightfield images of wildtype or homozygous hspb7 mutant 24 hpf embryos. (F) Representative 72 hpf hearts from wildtype or hspb7 mutant lines crossed onto a tg(myl7:gfp) background. (G) Quantification of normal hearts at 72 hpf in wildtype or homozygous hspb7 mutant animals (n = 200).