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Fig. 1 Generation of leap2-deficient zebrafish. (A) Phylogenetic tree of LEAP2. (B) Multiple sequence alignment columns with no gaps are colored in blue or red of LEAP2 precursor. The red color indicates highly conserved columns and blue indicates less conserved ones. (C) Alignment of mature LEAP2, complete conservation is marked with "?", molecules with roughly similar sizes and the same hydrophilicity and hydrophobicity are marked with ":", and residues whose molecular size and hydrophobicity are preserved to a certain extent are marked with "." (D) Relative expression levels of leap2 in various stages of zebrafish embryos. (E) Specific guide RNA (sgRNA) site is targeted in the first exon of leap2. (F) Electropherogram of Sanger DNA sequencing for leap2+/+ and leap2?/? zebrafish. (G) The gel electrophoresis of PCR products spanning mutated sites from mutant (?/?), wild type (+/+) and heterozygous (±). (H) The PCR products of leap2+/+ and leap2?/? zebrafish were digested by MmeI. (I) Comparison of DNA sequences identified in leap2+/+ and leap2?/? zebrafish lines. (J) The alignment of predicted protein sequences in leap2+/+ and l leap2?/? zebrafish lines. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)