Fig. 3.

Generation of flrt2-KO zebrafish line by CRISPR/Cas9 genome editing. (A) Sequencing maps of WT, flrt2^+/− mutants and flrt2^−/− mutants. The sequences in the red frame represent the areas targeted by CRISPR/Cas9. Blue background indicates CGG base pairs in WT and heterozygous zebrafish. (B) Sequence alignment of the WT and flrt2-KO zebrafish line, including the 20-bp deletion in homozygotes. (C) Reduced expression of flrt2 mRNA in the brain of WT and flrt2^−/− larvae (24 hpf) zebrafish analyzed by RT-qPCR (n=30/each group); t=23.58, P<0.0001. The solid bars represent the means±standard deviations. (D) Schematic diagram of FLRT2 protein translation. The yellow arrows indicate the process and direction of flrt2 mRNA translation into protein. Blue background means the position of the premature termination of protein translation. And the bottom indicates that the FLRT2 protein translation is forced to prematurely terminate the site.