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ZFIN ID: ZDB-FIG-170922-50
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:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Long-pec to Day 4
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Long-pec to Day 4

Fig. 3

Vascular ECs Are Required for HSPC Lodgment and Proliferation

(A) High-resolution confocal imaging of Tg (kdrl:mCherry/runx1:en-GFP) of control and klf6a morphants. White circles mark “cuddling HSPC.” Blue arrows indicate the localization and division of HSPCs associated with the CHT niche in control embryos. White arrows indicate HSPCs that could not seed the perivascular niche and migrated away from ECs in klf6a morphants. White arrowheads mark HSPCs without division in klf6a morphants. Scale bar, 30 μm.

(B) Quantification of lodging HSPCs in the CHT in control embryos and klf6a morphants at 51–53 hpf (mean ± SD, t test; p < 0.05, ∗∗p < 0.01, n = 6).

(C) Confocal imaging of Tg (kdrl:mCherry/CD41:GFP) at 3 dpf in control embryos and klf6a morphants. The left panel is high-resolution imaging showing dividing HSPCs. White circles mark “cuddling HSPCs.” White arrowheads indicate the HSPCs. Scale bars, 10 μm (left) and 50 μm (right).

(D) Confocal imaging showing CD41+ cells in the CHT at 4 dpf. The white dashed lines mark HSPCs and hematopoietic differentiated cells. Scale bar, 50 μm.

(E) Quantification of CD41:GFP+ cells shown in (D) in control and klf6a morphants (mean ± SD, t test; ∗∗p < 0.01, n = 5).

(F) The imaging of FISH and anti-BrdU staining in control and klf6a morphants at 60 hpf. Red color indicates BrdU signal and green color indicates cmyb expressed cells. White arrowheads indicate the BrdU+/cmyb+ double-positive cells. Scale bar, 50 μm.

(G) The statistical data show the percentage of cmyb+BrdU+/cmyb+ cells in (F) (mean ± SD, t test; p < 0.05, n = 5).

(H) The imaging of FISH and anti-BrdU staining in sibling and klf6a−/− embryos at 55 hpf. Red color indicates BrdU signal and green color indicates cmyb expressed cells. White arrowheads indicate the BrdU+/cmyb+ double-positive cells. Scale bar, 50 μm.

(I) The statistical data show the percentage of cmyb+BrdU+/cmyb+ cells in (H) (mean ± SD, t test; p < 0.05, n = 4).

See also Figure S3; Movies S1 and S2.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP la2Tg control Day 4 hematopoietic multipotent progenitor cell IFL
la2Tg + MO1-klf6a standard conditions Day 4 hematopoietic multipotent progenitor cell IFL
la2Tg; s896Tg control Protruding-mouth hematopoietic multipotent progenitor cell IFL
la2Tg; s896Tg + MO1-klf6a standard conditions Protruding-mouth hematopoietic multipotent progenitor cell IFL
GFP ioz1Tg; s896Tg control Long-pec hematopoietic stem cell IFL
ioz1Tg; s896Tg + MO1-klf6a standard conditions Long-pec hematopoietic stem cell IFL
mCherry ioz1Tg; s896Tg control Long-pec endothelial cell IFL
ioz1Tg; s896Tg + MO1-klf6a standard conditions Long-pec endothelial cell IFL
la2Tg; s896Tg control Protruding-mouth endothelial cell IFL
la2Tg; s896Tg + MO1-klf6a standard conditions Protruding-mouth endothelial cell IFL
myb TU control Pec-fin hematopoietic stem cell ISH
TU + MO1-klf6a standard conditions Pec-fin hematopoietic stem cell ISH
WT standard conditions Long-pec hematopoietic stem cell ISH
klf6aioz102/ioz102 standard conditions Long-pec hematopoietic stem cell ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
TU + MO1-klf6a standard conditions Pec-fin caudal hematopoietic tissue hematopoietic stem cell proliferation decreased occurrence, abnormal
ioz1Tg; s896Tg + MO1-klf6a standard conditions Long-pec caudal hematopoietic tissue hematopoietic stem cell decreased amount, abnormal
klf6aioz102/ioz102 standard conditions Long-pec caudal hematopoietic tissue hematopoietic stem cell proliferation decreased occurrence, abnormal
la2Tg + MO1-klf6a standard conditions Day 4 caudal hematopoietic tissue hematopoietic multipotent progenitor cell decreased amount, abnormal
la2Tg; s896Tg + MO1-klf6a standard conditions Protruding-mouth caudal hematopoietic tissue hematopoietic multipotent progenitor cell decreased amount, 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