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ZFIN ID: ZDB-FIG-160926-27
Monteiro et al., 2016 - Transforming Growth Factor β Drives Hemogenic Endothelium Programming and the Transition to Hematopoietic Stem Cells. Developmental Cell   38(4):358-70 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: Prim-5 to Long-pec
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Prim-5 to Long-pec

Fig. 6

Jag1a Is Required Downstream of TGF² Signaling for HSC Specification

(A) Expression of arterial markers in jag1aMO is unaffected when compared to wild-type embryos.

(B) Expression of runx1, cmyb, and gfi1aa in wild-type and jag1a morphants (MO) at 28 hpf. All the markers analyzed are reduced or absent in jag1a morphants. Red arrowheads indicate the remaining gene expression in the floor of the DA of jag1a morphants.

(C) HSPCs (yellow arrowheads) are severely reduced in the CHT of itga2b:GFP;Kdrl:HsRas-mCherry transgenic embryos at 48 hpf injected with the jag1aMO. Itga2b:GFP+ cells, magenta; Kdrl-HsRas:mCherry+ cells, green.

(D) Overexpression of jag1a with a Kdrl:jag1-V5 construct partially rescues the loss of runx1 and cmyb in the floor of the DA. 15 pg of the construct was used for this experiment. The numbers of embryos are shown in each panel as the number of embryos with phenotype/total number analyzed.

(E) Quantitation of the rescue effect observed in (D).

See also Figure S6.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP la2Tg ; s843Tg standard conditions Long-pec ventral wall of dorsal aorta hematopoietic multipotent progenitor cell IFL
la2Tg ; s843Tg + MO2-jag1a standard conditions Long-pec ventral wall of dorsal aorta hematopoietic multipotent progenitor cell IFL
gfi1aa WT standard conditions Prim-5 ventral wall of dorsal aorta ISH
WT + MO2-jag1a standard conditions Prim-5 ventral wall of dorsal aorta ISH
hey2 WT standard conditions Prim-5 artery ISH
WT + MO2-jag1a standard conditions Prim-5 artery ISH
mCherry la2Tg ; s843Tg standard conditions Long-pec trunk vasculature IFL
Long-pec ventral wall of dorsal aorta hematopoietic multipotent progenitor cell IFL
la2Tg ; s843Tg + MO2-jag1a standard conditions Long-pec trunk vasculature IFL
Long-pec ventral wall of dorsal aorta hematopoietic multipotent progenitor cell IFL
myb WT standard conditions Prim-5 ventral wall of dorsal aorta ISH
Prim-15 ventral wall of dorsal aorta ISH
WT + MO1-tgfbr2b standard conditions Prim-15 ventral wall of dorsal aorta ISH
WT + MO2-jag1a standard conditions Prim-5 ventral wall of dorsal aorta ISH
notch3 WT standard conditions Prim-5 artery ISH
WT + MO2-jag1a standard conditions Prim-5 artery ISH
runx1 WT standard conditions Prim-5 ventral wall of dorsal aorta ISH
Prim-15 ventral wall of dorsal aorta ISH
WT + MO1-tgfbr2b standard conditions Prim-15 ventral wall of dorsal aorta ISH
WT + MO2-jag1a standard conditions Prim-5 ventral wall of dorsal aorta ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-tgfbr2b standard conditions Prim-15 ventral wall of dorsal aorta myb expression decreased amount, abnormal
Prim-15 ventral wall of dorsal aorta runx1 expression decreased amount, abnormal
WT + MO2-jag1a standard conditions Prim-5 ventral wall of dorsal aorta gfi1aa expression decreased amount, abnormal
Prim-5 ventral wall of dorsal aorta myb expression decreased amount, abnormal
Prim-5 ventral wall of dorsal aorta runx1 expression decreased amount, abnormal
la2Tg ; s843Tg + MO2-jag1a standard conditions Long-pec hematopoietic multipotent progenitor cell decreased amount, abnormal
Long-pec intermediate cell mass of mesoderm 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, 38(4), Monteiro, R., Pinheiro, P., Joseph, N., Peterkin, T., Koth, J., Repapi, E., Bonkhofer, F., Kirmizitas, A., Patient, R., Transforming Growth Factor β Drives Hemogenic Endothelium Programming and the Transition to Hematopoietic Stem Cells, 358-70, Copyright (2016) with permission from Elsevier. Full text @ Dev. Cell