ZFIN ID: ZDB-FIG-160926-19
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 Day 4
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Prim-5 to Day 4

Fig. 3

TGFβ1 and TGFβ3 Are Required for Specification of HSCs

(A) Expression of runx1 in wild-type, tgfb1, tgfb2, and tgfb3 morphants.

(B) Expression of kdrl in wild-type, tgfb1, tgfb2, and tgfb3 morphants.

(C) Expression of dll4 in wild-type, tgfb1, tgfb2, and tgfb3 morphants.

(D) Expression of dlC in wild-type, tgfb1, tgfb2, and tgfb3 morphants. All samples were analyzed at 28 hpf.

(E-G) Maximum projections of itga2b:GFP; Kdrl:HRas-mCherry transgenic embryos in (E) uninjected, (F) tgfb1 morphants, and (G) tgfb3 morphants at 48 hpf. The images show part of the trunk DA and white arrowheads denote itga2b:GFP+ (green), kdrl:HRas-mCherry+ (magenta) HSPCs.

(H) HSPCs counts in the entire trunk region of uninjected, tgfb1, and tgfb3 morphant itga2b:GFP; Kdrl:HRas-mCherry transgenic embryos at 48 hpf (p value is indicated on the graph, n = 10 for each of the conditions).

(I-K) Expression of rag1 in the thymus at 4 dpf (red arrowheads) in (I) wild-type, (J) tgfb1 morphants, and (K) tgfb3 morphants.

(L) Expression of efnB2a in wild-type, tgfb1, and tgfb3 morphants.

(M) Expression of gata2b in wild-type, tgfb1, and tgfb3 morphants. The numbers of embryos are shown in each panel as the number of embryos with phenotype/total number analyzed.

See also Figure S3.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
dlc WT standard conditions Prim-5 artery ISH
Prim-5 artery vascular endothelium ISH
WT + MO2-tgfb1b + MO5-tgfb1a standard conditions Prim-5 artery ISH
Prim-5 artery vascular endothelium ISH
WT + MO2-tgfb3 standard conditions Prim-5 artery ISH
Prim-5 artery vascular endothelium ISH
WT + MO3-tgfb2 standard conditions Prim-5 artery ISH
Prim-5 artery vascular endothelium ISH
dll4 WT standard conditions Prim-5 artery ISH
Prim-5 artery vascular endothelium ISH
WT + MO2-tgfb1b + MO5-tgfb1a standard conditions Prim-5 artery ISH
Prim-5 artery vascular endothelium ISH
WT + MO2-tgfb3 standard conditions Prim-5 artery ISH
Prim-5 artery vascular endothelium ISH
WT + MO3-tgfb2 standard conditions Prim-5 artery ISH
Prim-5 artery vascular endothelium ISH
efnb2a WT standard conditions Prim-5 artery ISH
WT + MO2-tgfb1b + MO5-tgfb1a standard conditions Prim-5 artery ISH
WT + MO2-tgfb3 standard conditions Prim-5 artery ISH
EGFP la2Tg ; s843Tg standard conditions Long-pec hematopoietic multipotent progenitor cell IFL
Long-pec vascular endothelium IFL
Long-pec ventral wall of dorsal aorta IFL
Long-pec ventral wall of dorsal aorta hematopoietic multipotent progenitor cell IFL
la2Tg ; s843Tg + MO2-tgfb1b + MO5-tgfb1a standard conditions Long-pec hematopoietic multipotent progenitor cell IFL
Long-pec vascular endothelium IFL
Long-pec ventral wall of dorsal aorta IFL
Long-pec ventral wall of dorsal aorta hematopoietic multipotent progenitor cell IFL
la2Tg ; s843Tg + MO2-tgfb3 standard conditions Long-pec hematopoietic multipotent progenitor cell IFL
Long-pec vascular endothelium IFL
Long-pec ventral wall of dorsal aorta IFL
Long-pec ventral wall of dorsal aorta hematopoietic multipotent progenitor cell IFL
gata2b WT standard conditions Prim-5 ventral wall of dorsal aorta ISH
WT + MO2-tgfb1b + MO5-tgfb1a standard conditions Prim-5 ventral wall of dorsal aorta ISH
WT + MO2-tgfb3 standard conditions Prim-5 ventral wall of dorsal aorta ISH
kdrl WT standard conditions Prim-5 vascular endothelium ISH
WT + MO2-tgfb1b + MO5-tgfb1a standard conditions Prim-5 vascular endothelium ISH
WT + MO2-tgfb3 standard conditions Prim-5 vascular endothelium ISH
WT + MO3-tgfb2 standard conditions Prim-5 vascular endothelium ISH
mCherry la2Tg ; s843Tg standard conditions Long-pec hematopoietic multipotent progenitor cell IFL
Long-pec ventral wall of dorsal aorta hematopoietic multipotent progenitor cell IFL
la2Tg ; s843Tg + MO2-tgfb1b + MO5-tgfb1a standard conditions Long-pec hematopoietic multipotent progenitor cell IFL
Long-pec ventral wall of dorsal aorta hematopoietic multipotent progenitor cell IFL
la2Tg ; s843Tg + MO2-tgfb3 standard conditions Long-pec hematopoietic multipotent progenitor cell IFL
Long-pec ventral wall of dorsal aorta hematopoietic multipotent progenitor cell IFL
rag1 WT standard conditions Day 4 thymus T cell ISH
WT + MO2-tgfb1b + MO5-tgfb1a standard conditions Day 4 Not Detected thymus T cell ISH
WT + MO2-tgfb3 standard conditions Day 4 Not Detected thymus T cell ISH
runx1 WT standard conditions Prim-5 ventral wall of dorsal aorta hematopoietic multipotent progenitor cell ISH
WT + MO2-tgfb1b + MO5-tgfb1a standard conditions Prim-5 ventral wall of dorsal aorta hematopoietic multipotent progenitor cell ISH
WT + MO2-tgfb3 standard conditions Prim-5 ventral wall of dorsal aorta hematopoietic multipotent progenitor cell ISH
WT + MO3-tgfb2 standard conditions Prim-5 ventral wall of dorsal aorta hematopoietic multipotent progenitor cell ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO2-tgfb1b + MO5-tgfb1a standard conditions Prim-5 ventral wall of dorsal aorta gata2b expression decreased amount, abnormal
Prim-5 ventral wall of dorsal aorta hematopoietic multipotent progenitor cell runx1 expression decreased amount, abnormal
Day 4 thymus T cell rag1 expression absent, abnormal
WT + MO2-tgfb3 standard conditions Prim-5 ventral wall of dorsal aorta gata2b expression decreased amount, abnormal
Prim-5 ventral wall of dorsal aorta hematopoietic multipotent progenitor cell runx1 expression decreased amount, abnormal
Day 4 thymus T cell rag1 expression absent, abnormal
la2Tg ; s843Tg + MO2-tgfb1b + MO5-tgfb1a standard conditions Long-pec ventral wall of dorsal aorta hematopoietic multipotent progenitor cell decreased amount, abnormal
la2Tg ; s843Tg + MO2-tgfb3 standard conditions Long-pec ventral wall of dorsal aorta 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