Figure 1—figure supplement 1.

Figure 1—figure supplement 1. Zebrafish Vsx1 and Vsx2 proteins are disrupted in vsxKO animals.

(a, b). Scheme of zebrafish Vsx1 and Vsx2 peptides and protein alignments showing the nuclear localization signal (N, yellow), homeodomain (HD, blue) and CVC (green) regions. The starting of the mutation induced by CRISPR/Cas9 is indicated by a purple arrowhead. (a). The in-frame 53 amino acid deleted region in the vsx1 mutant is depicted by a purple color in the upper scheme and in the bottom protein alignment. (b). The 24 amino acid deleted region in the vsx2 mutant is depicted by a purple color in the upper scheme followed by a graded grey box indicating the frame-shift in the sequence and the premature stop codon generated in the CVC domain. Important to note is that the deletion in vsx2 comprised conserved critical amino acids that have been identified as causative mutations both in mice (ocular retardation model) and microphthalmic patients (R200Q/P and R227W mutations). (c). Western blot representative image using an anti-Vsx1 antibody showing the WT Vsx1 protein (black arrow). Note that Vsx1 protein is not detected either in vsxKO at 24hpf (middle lane) or in early (1.5hpf) wildtype embryos (right lane). (d). Western blot representative image using an anti-Vsx2 antibody showing the WT Vsx2 protein at 24hpf (black arrow). No Vsx2 protein is detectable in vsxKO heads at 24hpf.