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Fig. 1 Characterization of grhl3−/−(+14bp) mutants. (A) Three distinct phenotypes of grhl3−/−(+14bp) embryos are present at 72 hpf (classified as class 1, class 2, and class 3 based on the severity and incidence of phenotypes). (B) Structure of a functional Grhl protein comprising 557 amino acids, showing transactivation domain (orange), DNA binding domain (blue), and dimerization domain (yellow) relative to predicted non-functional proteins produced following CRISPR/Cas 9-mediated deletion in grhl3−/−(+14bp), grhl3−/−(−31bp), and grhl3−/−(Δ1-4) zebrafish. The grhl3 mutants were generated by targeting exon 4 at a site lying 3′ to seven putative in-frame ATG-initiation codons (indicated by red arrow) via CRISPR/Cas 9 mediated deletion and homologous integration. (C) The three distinct phenotypes (class1-3) of grhl3−/−(−31bp) and grhl3−/−(Δ1-4) embryos at 72 hpf. Scale bars: 200 μm.