Supplementary Figure 7: NC are required for HSCs; NC mutant and morphant validation.

Lateral trunk views (dorsal up, anterior left) of in situs for the NC marker crestin (a–h) and the HSC marker runx1 (i–p) at 24 h.p.f. in uninjected (a, i), Tfap2a morphant (b,j), Sdc4 morphant (c,k), Sox10 morphant (d,l), wild-type and sox10^m241 heterozygous (e,m), sox10^m241 homozygous mutant (f,n), wild-type and tfap2a^ts213 heterozygous (g,o) and sox10^ts213 homozygous mutant (h,p) embryos. Red brackets enclose the AGM region (a–h), green arrowheads indicate runx1 expressing cells (i–p). Numbers in the bottom right corner of panels indicate numbers of zebrafish embryos analysed for the indicated gene, versus the total number of zebrafish analysed. Zebrafish randomly selected from three clutches are represented by the image (a–p). Scale bars = 100 μm (a–p). Lateral images (dorsal up, anterior left) of whole uninjected (q), Tfap2a morphant (r), Sdc4 morphant (s) and Sox10 morphant (t) embryos at 24 h.p.f. showing embryo morphology. RT-PCR showing aberrant mRNA splicing in Tfap2a morphant (arrows) compared with uninjected embryos (u). Lateral views (dorsal up, anterior left) of uninjected controls (v), wild-type and sox10^m241 heterozygous (w), wild-type and tfap2a^ts213 heterozygous (x) embryos have increased numbers of melanophores compared with Sox10 morphant (v’), sox10^m241 homozygous mutant (w’) and tfap2a^ts213 homozygous mutant (x’) embryos (red arrowheads indicate presence of increased numbers of melanophores) at the developmental stages indicated (v–x’). Scale bars = 200 μm (q–t, v–y’).