Fig. 1.

Generation and characterization of ankrd26^ku6 zebrafish. (A) Schematic representation of human, mouse and zebrafish Ankrd26 protein, comprising N-terminal ankyrin (ANK) modules and C-terminal coiled-coil domain. (B) Schematic representation of the genomic DNA structure of ankrd26 and its encoded mRNA, showing the targeted area in the 5′-untranslated region (5′-UTR) of ankrd26 by CRISPR/Cas9 and the two deleted nucleotides (nt) in the 5′-UTR (Δ2). ORF, open reading frame. (C) Sanger sequencing confirmed the deletion of a two-nucleotide (GC) sequence (boxed). This mutant allele was named as ku6 in ZFIN. (D) Relative ankrd26 mRNA levels in pooled wild-type (wt), ankrd26^ku6/+ (heterozygote) and ankrd26^ku6 (homozygote) zebrafish larvae (n=10) at 5 days post-fertilization (dpf). (E,F) Western blotting (E) and densitometric analysis (F) of Ankrd26 protein expression levels in wt, ankrd26^ku6/+ and ankrd26^ku6 zebrafish larvae. β-Actin protein was used as protein loading control in E. Data are means±s.e.m. of at least three independent experiments. Mann–Whitney U-test was used to analyze differences between two groups. *P<0.05, **P<0.01 and ***P<0.005.