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ZFIN ID: ZDB-FIG-150323-3
Sawamiphak et al., 2014 - Interferon Gamma Signaling Positively Regulates Hematopoietic Stem Cell Emergence. Developmental Cell   31:640-653 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Condition:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Prim-5 to Day 4
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage Range: Prim-15 to High-pec

Fig. 3 Ifng1-2 Signaling Positively Regulates HSC Emergence

(A–F) Higher frequency of ECs at the aortic floor that turn on HSC marker expression upon ifng1-2 overexpression. Still images captured at different time points during time-lapse imaging of control (lacking the hsp:ifng1-2-V5 transgene) (A–C) and tg(hsp:ifng1-2-V5) (D–F) embryos starting at 33 hpf, 8 hr after heat shock (see also Movies S1 and S2). White arrowheads point to HSCs, labeled by Tg(itga2b:EGFP) (green) and Tg(kdrl:HRAS-mCherry) (red) expression, just arisen from the aortic wall at each indicated time point.

(G–L) Ifng1-2 knockdown reduces the emergence of HSCs and consequently the development of myeloid and lymphoid cells. runx1 expression was assessed during initiation of definitive HSC development at 26 hpf in mock-injected (G) and ifng1-2 MO-injected (H) embryos. Myeloid marker mpx1 expression in 48 hpf mock-injected (I) and ifng1-2 MO-injected (J) embryos. Lymphoid marker rag1 expression in the developing thymus (black circle) of mock-injected (K) and ifng1-2 MO-injected (L) 4 dpf larvae.

(M–P) Elevated expression of runx1 upon ifng1-2 overexpression. runx1 expression in 36-38 hpf WT embryos heat-shocked (h.s.) at 10–12 hpf (M) or 20–22 hpf (O) and Tg(hsp:ifng1-2) embryos heat-shocked at 10–12 hpf (N) or 20–22 hpf (P). The number of embryos showing the representative phenotype per total number of embryos analyzed is indicated in the lower left corner. Red brackets identify the dorsal aorta (DA), and blue brackets identify the cardinal vein (CV). All images are lateral views, dorsal up and anterior to the left.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP la2Tg; s896Tg control Prim-15 to High-pec hematopoietic stem cell IHC
Prim-15 to High-pec ventral wall of dorsal aorta hematopoietic stem cell IHC
la2Tg; s896Tg; s994Tg heat shock Prim-15 to High-pec hematopoietic stem cell IHC
Prim-15 to High-pec ventral wall of dorsal aorta hematopoietic stem cell IHC
mCherry la2Tg; s896Tg control Prim-15 to High-pec vasculature IHC
Prim-15 to High-pec ventral wall of dorsal aorta hematopoietic stem cell IHC
s896Tg; s994Tg heat shock Prim-15 to High-pec vasculature IHC
Prim-15 to High-pec ventral wall of dorsal aorta hematopoietic stem cell IHC
mpx WT control Long-pec myeloid cell ISH
WT + MO1-ifng1 standard conditions Long-pec myeloid cell ISH
rag1 WT control Day 4 common lymphoid progenitor ISH
Day 4 thymus ISH
WT + MO1-ifng1 standard conditions Day 4 common lymphoid progenitor ISH
Day 4 thymus ISH
runx1 WT control Prim-5 angioblastic mesenchymal cell ISH
Prim-25 angioblastic mesenchymal cell ISH
Prim-25 hematopoietic stem cell ISH
WT + MO1-ifng1 standard conditions Prim-5 angioblastic mesenchymal cell ISH
s994Tg heat shock Prim-25 angioblastic mesenchymal cell ISH
Prim-25 hematopoietic stem cell ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
s896Tg; s994Tg heat shock Prim-15 - High-pec endothelial to hematopoietic transition increased process quality, 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, 31, Sawamiphak, S., Kontarakis, Z., Stainier, D.Y., Interferon Gamma Signaling Positively Regulates Hematopoietic Stem Cell Emergence, 640-653, Copyright (2014) with permission from Elsevier. Full text @ Dev. Cell