ZFIN ID: ZDB-FIG-190910-29
Bonkhofer et al., 2019 - Blood stem cell-forming haemogenic endothelium in zebrafish derives from arterial endothelium. Nature communications   10:3577 Full text @ Nat. Commun.
Anatomical Terms:
Stage: Prim-5

Fig. 2

FACS based isolation of haemogenic and aortic roof endothelium. a Schematic of transgene expression in the trunk region of double transgenic TgBAC(runx1P2:Citrine);Tg(kdrl:mCherry) embryos during definitive haematopoiesis. b Representative confocal image of the DA region of double transgenic TgBAC(runx1P2:Citrine);Tg(kdrl:mCherry) embryos at 29 hpf. Laser intensities enhanced to detect Citrine fluorescence in the DA roof and sprouting inter-somitic vessels (ISV) (orange arrows) in addition to the HE (green arrow). Yellow arrow heads: neurons in the spinal cord. c Characteristic FACS plot of the established gating strategy for the isolation of endothelial sub-populations (runx1hikdrl+ [DP-R1hi]; runx1medkdrl+ [DP-R1med]; runx1lokdrl+[DP-R1lo]; runx1kdrl+ [SP-kdrl]) from double transgenic TgBAC(runx1P2:Citrine);Tg(kdrl:mCherry) embryos. d qRT-PCR analysis of haematopoietic (tal1cmyb and gfi1aa) and endothelial (tbx20efnb2a and dll4) marker gene expression in the different cell fractions isolated from ~29 hpf embryos following the FACS gating strategy depicted in (c). Graphs show the measured mean fold change relative to the expression detected in the DP-R1lo fraction. n = 4 independent experiments. Error bars represent the SEM. 1-way ANOVA; *p < 0.05; **p < 0.01; ***p < 0.001. e Schematic of the vessel structure in the zebrafish trunk region discriminating between aortic roof endothelium (ARE) including the ISV, the HE and the non-aortic endothelium (NAE) including but not restricted to endothelial cells from the cardinal vein (CV). The depicted colour code refers to the colours of the FACS gates presented in (c)

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
Citrine ox1Tg; s896Tg standard conditions Prim-5 intersegmental vessel IFL
Prim-5 spinal cord neuron IFL
Prim-5 ventral wall of dorsal aorta IFL
Antibody Labeling Details No data available
Phenotype Details No data available
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Nat. Commun.