Extended Data Fig. 1 |. Phenotype characterization of zebrafish mutant^cas005.

a, Normal primitive haematopoiesis is intact in mut^cas005. WISH results demonstrate that the expression of primitive haematopoietic cell markers is identical between siblings and mut^cas005 embryos at 22 h.p.f., including scl (also known as tal1; haematopoietic progenitor marker), gata1 (also known as gata1a; erythrocyte progenitor marker) and pu.1 (also known as spi1b; myeloid progenitor marker). b, The vascular development is normal in mut^cas005 embryos. WISH results show no difference in the expression of kdrl (pan-endothelial cell marker) at 36 h.p.f. between wild-type siblings and mut^cas005 embryos. c, The haemogenic endothelium is intact in mut^cas005 embryos. WISH results show no difference in runx1 expression at 36 h.p.f. in wild-type and mut^cas005 embryos. d, e, The definitive haematopoiesis is defective in zebrafish mut^cas005 embryos. d, Bright-field images of wild-type and mut^cas005 embryos show no obvious morphological difference at 5 d.p.f. e, WISH results of myb, hbae1.1, mpx and lyz expression in wild-type and mut^cas005 embryos at 5 d.p.f. Arrows indicate the comparable position in wild-type (black) or mut^cas005 embryos (red). In 5 d.p.f. wild-type embryos, myb was expressed in all haematopoietic tissues including the CHT, thymus and kidney, whereas homozygous mut^cas005 embryos displayed markedly decreased myb expression in the CHT, but similar expression to wild-type embryos in the thymus and kidney marrow. In accordance, the expression of downstream haematopoietic lineage cell markers, including hbae1.1 (erythrocyte marker), mpx (granulocyte marker) and lyz (macrophage marker), also showed similar expression patterns in the wildtype (black) and mut^cas005 (red) embryos to that of the myb WISH analysis. f, The percentage of apoptotic HSPCs detected by the TUNEL assay in 2 and 4 d.p.f. wild-type and mut^cas005 embryos. 2 d.p.f.: P = 0.35, t = 0.96, df = 27; 4 d.p.f.: P = 0.50, t = 0.69, df = 27. Error bars denote s.e.m. g, At 7 d.p.f., myb expression in the thymus and kidney marrow was identical in wild-type sibling and mut^cas005 embryos, whereas mut^cas005 embryos still displayed markedly decreased myb expression in the CHT. In addition, mut^cas005 embryos displayed notably decreased hbae1.1, mpx and lyz expression in the CHT but not in the kidney marrow.