Fig. S1

hlx1 regulates hematopoietic stem cell specification and definitive myelopoiesis in zebrafish

(a) qPCR analysis for hlx1 expression in fli:hHLXOE endothelial cells and hlx1MO embryos (n=2: mean +s.d.). (b) TUNEL assay at 48hpf in hlx1MO or fli:hHLXOE embryos. The positive control was treated with DNase. Numbers in the bottom right corner of panels indicate numbers of zebrafish embryos with the indicated phenotype, versus the total number of zebrafish analyzed (n=3). (c) WISH for runx1 at 26hpf and c-myb at 72hpf, in control or Runx:hHLXOE embryos. Numbers in the bottom right corner as described above. (d) qPCR results for c-myb and terminal myeloid markers as indicated, in control and Runx:hHLXOE whole embryos at 5dpf. (n=2, mean +s.d.). (e) WISH for terminal myeloid markers as indicated, in control or Runx:hHLXOE embryos at 5dpf. Numbers in the bottom right corner as described above. (f) WISH for c-myb, in control or hlx1MO embryos at 36hpf. Numbers in the bottom right corner as described above. (g) Graphs depicting the percentage of Runx+ HSPCs in control Tg(Runx:mCherry) or hlx1MO injected with either a translational or a splicing morpholino at 48hpf measured by flow cytometry (n=3; mean + s.d.; Student’s t-test, *P<0.05). (h) WISH for ephrinB2α (arterial endothelium) and kdrl (vascular endothelium) in control or hlx1 MO embryos at 36hpf. Numbers in the bottom right corner as described above. (i) Representative images of Tg(Runx:mCherry) embryos at 72hpf, injected with hlx1 morpholino and the indicated amounts of Runx:hHLX construct. The numbers of mCherry-positive HSPCs from each embryo are represented in the graph (mean + s.d., ANOVA test, ***P<0.001).