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ZFIN ID: ZDB-FIG-160926-18
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 Reagent:
Anatomical Terms:
Stage Range: Prim-5 to Day 5
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Prim-5 to Day 5

Fig. 2

TGFβ Signaling Is Required for Specification of HSCs

Expression of runx1 in (A) wild-type (wt) and (B) tgfbr2 morphants at 26 hpf. Expression of gfi1aa in (C) wild-type or (D) tgfbr2 morphants at 26 hpf. Expression of gata2b in (E) wild-type or (F) tgfbr2 morphants at 26 hpf.

Expression of the vascular marker kdrl (G, H) and the arterial markers notch3 (I, J), hey2 (K, L), and efnB2A (M, N) is unaffected in tgfbR2 morphants. Maximum projections of itga2b:GFP; Kdrl:HRas-mCherry transgenic embryos at 48 hpf in (O) uninjected and (P) tgfbr2 MO1-injected embryos. Region shown includes part of the DA and white arrowheads denote itga2b:GFP+(green), kdrl:HRas-mCherry+ (magenta) HSPCs.

(Q) HSPC counts in the entire trunk region of uninjected and tgfbR2 morphant itga2b:GFP; Kdrl:HRas-mCherry transgenic embryos at 48 hpf (p value is indicated on the graph, n = 12 (wild-type) and n = 11 (tgfbR2MO1).

itga2b-GFP+ cells are present in the CHT of itga2b:GFP embryos (R, yellow arrowheads) and greatly reduced in the CHT of tgfbR2 morphants at 5 dpf (S). Expression of rag1 in the thymus (red arrowheads) at 4 dpf in (T) wild-type and (U) tgfbR2 morphants. Expression of hbbe1 in the CHT at 4 dpf in (V) wild-type and (W) tgfbR2 morphants.

The numbers of embryos are shown in each panel as the number of embryos with phenotype/total number analyzed. See also Figure S2.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
efnb2a WT standard conditions Prim-5 artery ISH
Prim-5 artery vascular endothelium ISH
WT + MO1-tgfbr2b standard conditions Prim-5 artery ISH
Prim-5 artery vascular endothelium ISH
EGFP la2Tg ; s843Tg standard conditions Long-pec hematopoietic multipotent progenitor cell IFL
Day 5 hematopoietic multipotent progenitor cell IFL
la2Tg ; s843Tg + MO1-tgfbr2b standard conditions Long-pec hematopoietic multipotent progenitor cell IFL
Day 5 hematopoietic multipotent progenitor cell IFL
gata2b WT standard conditions Prim-5 ventral wall of dorsal aorta ISH
WT + MO1-tgfbr2b standard conditions Prim-5 ventral wall of dorsal aorta ISH
gfi1aa WT standard conditions Prim-5 ventral wall of dorsal aorta ISH
WT + MO1-tgfbr2b standard conditions Prim-5 ventral wall of dorsal aorta ISH
hbbe1.1 WT standard conditions Day 4 erythroid lineage cell ISH
WT + MO1-tgfbr2b standard conditions Day 4 erythroid lineage cell ISH
hey2 WT standard conditions Prim-5 artery ISH
Prim-5 artery vascular endothelium ISH
WT + MO1-tgfbr2b standard conditions Prim-5 artery ISH
Prim-5 artery vascular endothelium ISH
kdrl WT standard conditions Prim-5 vascular endothelium ISH
WT + MO1-tgfbr2b standard conditions Prim-5 vascular endothelium ISH
mCherry la2Tg ; s843Tg standard conditions Long-pec dorsal aorta IFL
Long-pec hematopoietic multipotent progenitor cell IFL
Long-pec ventral wall of dorsal aorta IFL
la2Tg ; s843Tg + MO1-tgfbr2b standard conditions Long-pec dorsal aorta IFL
Long-pec hematopoietic multipotent progenitor cell IFL
Long-pec ventral wall of dorsal aorta IFL
notch3 WT standard conditions Prim-5 artery ISH
Prim-5 artery vascular endothelium ISH
WT + MO1-tgfbr2b standard conditions Prim-5 artery ISH
Prim-5 artery vascular endothelium ISH
rag1 WT standard conditions Day 4 thymus T cell ISH
WT + MO1-tgfbr2b standard conditions Day 4 Not Detected thymus T cell ISH
runx1 WT standard conditions Prim-5 ventral wall of dorsal aorta ISH
WT + MO1-tgfbr2b 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-5 ventral wall of dorsal aorta gata2b expression decreased amount, abnormal
Prim-5 ventral wall of dorsal aorta gfi1aa expression decreased amount, abnormal
Prim-5 ventral wall of dorsal aorta runx1 expression decreased amount, abnormal
Day 4 erythroid lineage cell hbbe1.1 expression decreased amount, abnormal
Day 4 thymus T cell rag1 expression absent, abnormal
la2Tg ; s843Tg + MO1-tgfbr2b standard conditions Long-pec hematopoietic multipotent progenitor cell decreased amount, abnormal
Long-pec ventral wall of dorsal aorta hematopoietic multipotent progenitor cell decreased amount, abnormal
Day 5 hematopoietic multipotent progenitor cell decreased amount, abnormal
Day 5 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