ZFIN ID: ZDB-FIG-170922-53
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 Reagents:
Anatomical Terms:
Stage Range: Long-pec to Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Long-pec to Protruding-mouth

Fig. 6

Ccl25b Acts Downstream of Klf6a to Facilitate HSPC Settlement and Expansion

(A) Time-lapse imaging of kdrl:mCherry/runx1:en-GFP in control and ccl25b morphants. The white arrowheads mark HSPCs and the white circles indicate HSPC pockets. Scale bar, 50 μm.

(B) The statistical data of runx1+ cells in D/M/V of CVP in control and ccl25b morphants at 50 hpf (mean ± SD, t test; p < 0.05, ns [not significant] > 0.05, n = 8).

(C) Confocal imaging of kdrl:mCherry/runx1:en-GFP CHT at 3 dpf in control embryos and ccl25b morphants. Yellow arrowheads mark GFP+ cells. The white dashed lines mark the outlines of caudal artery and vein. Scale bar, 50 μm.

(D) The quantification of runx1:en-GFP+ cells shown in (C) (mean ± SD, t test; ∗∗p < 0.01, n = 8).

(E) Confocal imaging of FISH (cmyb)/immunofluorescence (PH3) at 60 hpf in CHT of control embryos and ccl25b morphants. White arrowheads mark double-positive cells. Scale bar, 50 μm.

(F) The statistical data showing that the percentage of double-positive/cmyb+ cells was reduced significantly in the morphants (mean ± SD, t test; ∗∗∗p < 0.001, n = 8).

(G) qPCR result of cell division-related gene expression in control and ccl25b morphants (mean ± SD, t test; p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ns > 0.05, n = 3).

See also Figure S6.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
ccnd1 TU + MO2-ccl25b standard conditions Pec-fin to Protruding-mouth whole organism posterior region RTPCR
cdc5l TU + MO2-ccl25b standard conditions Pec-fin to Protruding-mouth whole organism posterior region RTPCR
cdk1 TU + MO2-ccl25b standard conditions Pec-fin to Protruding-mouth whole organism posterior region RTPCR
cdk2 TU + MO2-ccl25b standard conditions Pec-fin to Protruding-mouth whole organism posterior region RTPCR
cdkn1a TU + MO2-ccl25b standard conditions Pec-fin to Protruding-mouth whole organism posterior region RTPCR
GFP ioz1Tg; s896Tg control Long-pec hematopoietic stem cell IFL
Protruding-mouth hematopoietic stem cell IFL
ioz1Tg; s896Tg + MO1-ccl25b standard conditions Long-pec hematopoietic stem cell IFL
Protruding-mouth hematopoietic stem cell IFL
mCherry ioz1Tg; s896Tg control Long-pec endothelial cell IFL
Protruding-mouth endothelial cell IFL
ioz1Tg; s896Tg + MO1-ccl25b standard conditions Long-pec endothelial cell IFL
Protruding-mouth endothelial cell IFL
mnd1 TU + MO2-ccl25b standard conditions Pec-fin to Protruding-mouth whole organism posterior region RTPCR
myb TU standard conditions Pec-fin hematopoietic stem cell ISH
TU + MO2-ccl25b standard conditions Pec-fin hematopoietic stem cell ISH
ncapd2 TU + MO2-ccl25b standard conditions Pec-fin to Protruding-mouth whole organism posterior region RTPCR
nusap1 TU + MO2-ccl25b standard conditions Pec-fin to Protruding-mouth whole organism posterior region RTPCR
zw10 TU + MO2-ccl25b standard conditions Pec-fin to Protruding-mouth whole organism posterior region RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
TU + MO2-ccl25b standard conditions Pec-fin caudal hematopoietic tissue hematopoietic stem cell proliferation decreased occurrence, abnormal
TU + MO2-ccl25b standard conditions Pec-fin - Protruding-mouth whole organism posterior region cdk1 expression decreased amount, abnormal
TU + MO2-ccl25b standard conditions Pec-fin - Protruding-mouth whole organism posterior region cdc5l expression decreased amount, abnormal
TU + MO2-ccl25b standard conditions Pec-fin - Protruding-mouth whole organism posterior region zw10 expression decreased amount, abnormal
TU + MO2-ccl25b standard conditions Pec-fin - Protruding-mouth whole organism posterior region ccnd1 expression decreased amount, abnormal
TU + MO2-ccl25b standard conditions Pec-fin - Protruding-mouth whole organism posterior region cdk2 expression decreased amount, abnormal
TU + MO2-ccl25b standard conditions Pec-fin - Protruding-mouth whole organism posterior region ncapd2 expression decreased amount, abnormal
TU + MO2-ccl25b standard conditions Pec-fin - Protruding-mouth whole organism posterior region nusap1 expression decreased amount, abnormal
TU + MO2-ccl25b standard conditions Pec-fin - Protruding-mouth whole organism posterior region mnd1 expression decreased amount, abnormal
TU + MO2-ccl25b standard conditions Pec-fin - Protruding-mouth whole organism posterior region cdkn1a expression increased amount, abnormal
ioz1Tg; s896Tg + MO1-ccl25b standard conditions Long-pec hematopoietic stem cell migration decreased occurrence, abnormal
Protruding-mouth caudal hematopoietic tissue hematopoietic stem cell proliferation decreased occurrence, 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