Fig. 6

Bonkhofer et al., 2019 - Blood stem cell-forming haemogenic endothelium in zebrafish derives from arterial endothelium
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Fig. 6

Aortic HE maintains arterial features in the absence of runx1. a qRT-PCR gene expression analysis of the arterial marker efnb2aand the venous marker ephb4b in the HE and ARE of runx1+/+ (WT) and the HE of runx1−/− mutant (MUT) embryos. Graphs show the mean of detected expression levels relative to the geometric mean (GM) of the two housekeeping genes eef1a1l and rpl13an = 5 independent biological experiments for WT embryos and n = 6 independent biological experiments for MUT embryos. Error bars represent the SEM. Two-way ANOVA; *p < 0.05; **p < 0.01; ***p < 0.001. b Heatmaps of whole-genome gene expression data from RNA-seq for the HE of control and runx1 MO embryos showing arterial and venous genes. cRepresentative fluorescent microscopy image of 6 dpf TgBAC(runx1P2:Citrine), Tg(kdrl:mCherry) double transgenic embryos on a runx1+/+ or runx1−/− genetic background. Left: Region depicting the region of definitive haematopoietic niches including the thymus (Th) and the kidney (Ki). Right: region depicting the DA and the beginning of the caudal haematopoietic tissue (CHT). Green arrows point to the ventral wall of the DA. d Quantification of embryos derived from the indicated in-crosses (InX) depicting the runx1−/− mutant phenotype as depicted in (c). e ISH analysis of runx1 in control and dll4 MO embryos. Experiment was performed with two different MOs. Green arrows point to the HE. f Offspring of dll4+/− heterozygous fish were analysed by ISH for runx1 at 32 hpf. Runx1 expression was quantified by image analysis and subsequent genotyping57. Representative examples of runx1 expression in each of the WT, dll4+/− and dll4−/− genotypes are shown in the right-hand panels and the corresponding expression values are shown as orange points in the graph. (μwt = 33.3, μmut = 15.19, p < 0.0001, F(2,50) = 16,79, one way ANOVA). g Model depicting the lineage relationship between the definitive HE giving rise to HSCs and arterial endothelium in the DA during embryogenesis. Dll4+ aortic progenitors get exposed to localised signals patterning the DA and inducing haemogenic gene expression. Haemogenic specification includes upregulation of runx1 expression and a subsequent Runx1 induced repression of dll4, thus giving rise to Dll4 mature HE

Expression Data
Knockdown Reagents:
Anatomical Terms:
Stage Range: Prim-15 to Day 6

Expression Detail
Antibody Labeling
Phenotype Data
Knockdown Reagents:
Observed In:
Stage Range: Prim-15 to Day 6

Phenotype Detail
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