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ZFIN ID: ZDB-FIG-170922-49
Xue et al., 2017 - The Vascular Niche Regulates Hematopoietic Stem and Progenitor Cell Lodgment and Expansion via klf6a-ccl25b. Developmental Cell   42(4):349-362.e4 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Anatomical Terms:
Stage Range: Prim-15 to Day 5
PHENOTYPE:
Fish:
Observed In:
Stage Range: Long-pec to Day 5

Fig. 2

The klf6a-Deficient Embryos Display HSPC Defects in the CHT

(A) The high-resolution expression pattern of klf6a in the CHT at 33 hpf and 48 hpf. Black arrowheads indicate the expression of klf6a in caudal venous ECs. Scale bar, 30 μm.

(B) Double FISH shows klf6a (red) co-localized with fli1a (green) at 33 hpf in the CHT. White arrowheads indicate the expression of klf6a, fli1a or the co-expression of fli1a and klf6a, respectively. Scale bar, 30 μm.

(C) The mutation of klf6a with a 7-bp deletion in genomic DNA and the predicated truncated protein.

(D) Western blot result of Klf6a in the siblings and mutants at 48 hpf.

(E) Confocal imaging showing kdrl+ vascular deficiency in the CHT and head region, whereas gata1+ cells accumulated in the brain in klf6a−/− embryos in Tg (kdrl:GFP/gata1:dsRed) background. Scale bar, 50 μm.

(F) 3D imaging of the CHT vascular structure. Scale bar, 50 μm.

(G) Confocal imaging of runx1+ cells and vascular structure in the siblings and klf6a/ embryos in Tg (kdrl:mCherrry/runx1:en-GPF) background at 60 hpf and 4 dpf (mean ± SD, t test; p < 0.05, ns [not significant] > 0.05, n = 5). Scale bar, 50 μm.

(H) The qPCR results of cmyb, pu.1, and ae1-globin expression in siblings and klf6a/ embryos at 5 dpf (mean ± SD, t test; p < 0.05, n = 3).

(I) Expression of cmyb, lyz, and gata1 in siblings and klf6a/ embryos at 4 dpf by WISH. Black arrowheads indicate the expression pattern of these markers.

See also Figure S2.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
DsRed s843Tg ; sd2Tg ; klf6aioz102/ioz102 standard conditions Long-pec hematopoietic cell IFL
s843Tg; sd2Tg standard conditions Long-pec hematopoietic cell IFL
EGFP s843Tg ; sd2Tg ; klf6aioz102/ioz102 standard conditions Long-pec caudal vein plexus IFL
Long-pec vasculature IFL
s843Tg; sd2Tg standard conditions Long-pec caudal vein plexus IFL
Long-pec vasculature IFL
fli1a TU standard conditions Prim-15 endothelial cell ISH
gata1a WT standard conditions Day 4 hematopoietic cell ISH
klf6aioz102/ioz102 standard conditions Day 4 hematopoietic stem cell ISH
GFP ioz1Tg ; s896Tg ; klf6aioz102/ioz102 standard conditions Pec-fin caudal vein plexus hematopoietic stem cell IFL
Day 4 caudal vein plexus hematopoietic stem cell IFL
ioz1Tg; s896Tg standard conditions Pec-fin caudal vein plexus hematopoietic stem cell IFL
Day 4 caudal vein plexus hematopoietic stem cell IFL
hbae1.1 klf6aioz102/ioz102 standard conditions Day 5 whole organism posterior region RTPCR
klf6a TU standard conditions Prim-15 endothelial cell ISH
Prim-15 intermediate cell mass of mesoderm posterior region ISH
Long-pec intermediate cell mass of mesoderm posterior region ISH
Ab1-klf6a WT standard conditions Long-pec whole organism WB
Ab1-klf6a klf6aioz102/ioz102 standard conditions Long-pec whole organism WB
lyz WT standard conditions Day 4 hematopoietic cell ISH
klf6aioz102/ioz102 standard conditions Day 4 hematopoietic stem cell ISH
mCherry ioz1Tg ; s896Tg ; klf6aioz102/ioz102 standard conditions Pec-fin caudal vein plexus IFL
Day 4 caudal vein plexus IFL
ioz1Tg; s896Tg standard conditions Pec-fin caudal vein plexus IFL
Day 4 caudal vein plexus IFL
myb WT standard conditions Day 4 hematopoietic cell ISH
klf6aioz102/ioz102 standard conditions Day 4 hematopoietic stem cell ISH
Day 5 whole organism posterior region RTPCR
spi1b klf6aioz102/ioz102 standard conditions Day 5 whole organism posterior region RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
ioz1Tg ; s896Tg ; klf6aioz102/ioz102 standard conditions Pec-fin caudal vein plexus hematopoietic stem cell decreased amount, abnormal
Pec-fin hematopoietic stem cell migration decreased occurrence, abnormal
Day 4 caudal vein plexus hematopoietic stem cell decreased amount, abnormal
Day 4 hematopoietic stem cell migration decreased occurrence, abnormal
klf6aioz102/ioz102 standard conditions Long-pec whole organism klf6a expression decreased amount, abnormal
Day 4 hematopoietic stem cell lyz expression decreased amount, abnormal
Day 4 hematopoietic stem cell myb expression decreased amount, abnormal
Day 4 hematopoietic stem cell gata1a expression decreased amount, abnormal
Day 4 hematopoietic stem cell gata1a expression decreased distribution, abnormal
Day 4 hematopoietic stem cell myb expression decreased distribution, abnormal
Day 4 hematopoietic stem cell lyz expression decreased distribution, abnormal
Day 5 whole organism posterior region myb expression decreased amount, abnormal
Day 5 whole organism posterior region spi1b expression decreased amount, abnormal
Day 5 whole organism posterior region hbae1.1 expression decreased amount, abnormal
s843Tg ; sd2Tg ; klf6aioz102/ioz102 standard conditions Long-pec brain vasculature hemorrhagic, abnormal
Long-pec caudal vein plexus decreased thickness, abnormal
Long-pec caudal vein plexus decreased volume, abnormal
Long-pec caudal vein plexus disorganized, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Cell for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Developmental Cell, 42(4), Xue, Y., Lv, J., Zhang, C., Wang, L., Ma, D., Liu, F., The Vascular Niche Regulates Hematopoietic Stem and Progenitor Cell Lodgment and Expansion via klf6a-ccl25b, 349-362.e4, Copyright (2017) with permission from Elsevier. Full text @ Dev. Cell