Supplementary Figure 3: Pdgfr-mediated signalling is required for NC migration to the DA.

NC migration visualized by crestin expression in lateral (dorsal up, anterior left) and transverse (dorsal up) views of DMSO (a–a’) or inhV (b–b’) treated embryos, uninjected embryos (c-c’), a subset of embryos injected with 10 ng of PdgfrbMO displaying normal NC migration (d–d’) a subset of embryos injected with 10 ng of PdgfrbMO displaying defective NC migration (e–e’) and embryos injected with both PdgfraMO and PdgfrbMO (f–f’). 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–f,h,i). Positions of transverse sections are indicated by the red bars (a–f). Scale bars = 100 μm (a–f, h-i). Average number of NC streams ventral to the notochord at 24 h.p.f. for the indicated conditions, error bars indicate standard error of the mean, n = 20 is the number of randomly selected zebrafish embryos from three biologically independent clutches each, the triple asterisk indicates a statistically significant result between bars, from left to right, p-value was < 0.0001 by Student’s t-test in all cases (g), see Supplementary Table 3. Lateral views (dorsal up, anterior left) of crestin expression in uninjected (h) and Pdgfra morphant (i) embryos at 36 h.p.f. NC streams that have migrated ventral to the notochord are indicated by yellow arrowheads (a–f,h–i), red brackets enclose the AGM region (a–f), green arrowheads indicate the presence of melanophores (h–i). Lateral z-stack volume projections of the trunk (dorsal up, anterior left) with vasculature in green (fli1a:EGFP) and NC in red (sox10:mRFP) in uninjected (j) and Pdgfra morphant (k) embryos, blue (k) and yellow (j) arrowheads indicate NC streams which are dorsal to and ventral to the notochord respectively, scale bar = 100 μm, mag. = 200X. nt – neural tube, no – notochord, DA – dorsal aorta, PCV – posterior cardinal vein.