Figure 1. Establishment of Gli3 mutant cell lines.

(A) Exon-intron structure of the mouse Gli3 gene. Exon2 contains the translation initiation codon. (B) A part of the Gli3 sequences with the target sites of DSBs (arrows) by Cas9 nuclease. Target sequences for sgRNA are underlined, and PAMs are shown in red. Numerals over the WT sequence indicate the nucleotide position in the ORF sequence from the first base of the start codon (+1). Identical clones are indicated by the colored allele names in parentheses. The six clones with bold letters having biallelic out-of-frame mutations were subjected to Western analysis.