Supplementary Figure 6: NC are required for HSCs.
From: Pdgf signalling guides neural crest contribution to the haematopoietic stem cell specification niche
Lateral trunk views (dorsal up anterior left) of in situs for the HSC marker runx1(blue) and the NC marker crestin (red) (a,d,g,j,m), the HSC marker cmyb (b,e,h,k,n), and the T lymphocyte marker rag1 (c,f,i,l,o, ventral head views) in uninjected or Tfap2a morphant (a–f), wild-type and sox10m241heterozygous (g–i), sox10m241 homozygous mutant (j–l) and Sdc4 morphant (m–o) embryos at the developmental stages indicated. Green and red arrowheads (a,b,d,e,g,h,j,k,m,n) indicate cells expressing runx1 and cmyb respectively, blue arrowheads (c,f,i,l,o) identify rag1 expressing T-lymphocytes when present, red brackets (a,d,g,j,m) enclose the AGM region. Numbers in bottom right corner indicate number of individuals in the clutches represented by the image (a–o). Scale bars = 100 μm (a–o). Average number of runx1+ (p) or cmyb+ (q) DA cells, or average number of NC streams ventral to the notochord at 24 h.p.f (r) in the indicated conditions. 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–o). Fluorescently labelled HSCs in uninjected (s) and Sdc4 morphant (t) cmyb:GFP;kdrl:mCherry transgenic embryos at 36 h.p.f., HSCs are yellow cells indicated by yellow arrowheads, scale bar = 50 μm, mag. = 200X. Average number of mCherry+GFP+ cells in the DA for the indicated conditions (u), n = 8 zebrafish each. Error bars indicate the standard error of the mean (p–r,u). Triple asterisk indicates p-values < 0.0001 in (p–r) and p = 0.0007 in (u) by Student’s t-test. See Supplementary Table 3.
