Fig. S5

MZfscn1a mutants display migration defects in the first NC stream.

(a) Generation of fscn1a mutant with TALENs. The “Left” and “Right” binding sites for TALENs were underlined and the BSRI site was shown in green used for detection of mutations by restriction enzyme digestion. One mutation which led to a shift of the open reading frame with a premature stop codon (red) was identified from the F1 embryos of founder fish. (b-c) The expression of fscn1a mRNA and protein in wild-type and MZfscn1a mutant embryos. Wild-type and MZfscn1a mutant embryos were harvested at the 75%-epiboly stage for in situ hybridization (b) and immunoblotting (c). Note that the forerunner cell expression of fscn1a transcripts is not changed in MZfscn1a mutants (b, indicated by arrow head). Scale bar, 200 µm. (d) Wild-type and MZfscn1a mutant embryos were harvested at 11 hpf (Scale bar, 200 µm), 18 hpf (Scale bar, 100 µm) and 28 hpf (Scale bar, 200 µm) for in situ hybridization with foxd3 and dlx2a probes. Note that MZfscn1a mutants have normal NC induction but decreased number of dlx2a-expressing NC cells in the first stream.