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ZFIN ID: ZDB-FIG-150323-5
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 Reagents:
Anatomical Terms:
Stage Range: Prim-5 to Long-pec
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage: Long-pec

Fig. 5

Reduced Ifng1-2 Signaling Is Partially Responsible for the HSC Developmental Defects Caused by the Lack of Blood Flow

(A–D) ifng1-2 overexpression partly restores HSCs in embryos with no circulation. itga2b:EGFP+ (green) kdrl:HRAS-mCherry+ (red) HSCs (white arrowheads) are shown in mock-injected controls (A) and tnnt2 MO-injected embryos without (B) and with (C) hsp:ifng1-2 transgene. All embryos were heat shocked at 24 and 48 hpf and imaged at 52–54 hpf. (D) HSC numbers per 500 µm aortic length are shown as means ± SEM, n = 12–20 embryos, p d 0.01, p d 0.001.

(E–G) Loss of runx1 expression from the lack of blood flow is rescued by ifng1-2 overexpression. runx1 expression is shown in mock-injected controls (E) and tnnt2 MO-injected embryos without (F) and with (G) the hsp:ifng1-2 transgene. All embryos were heat-shocked at 24 hpf and harvested at 36 hpf.

(H–K) crfb17 and ifng1-2 expression depends on blood flow. crfb17 (H and I) and ifng1-2 (J and K) expression is shown in mock-injected (H and J) and tnnt2 MO-injected (I and K) 36 hpf embryos. 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. qPCR assay of ifng1-2 and crfb17 expression at 36 hpf in tnnt2 MO-injected embryos compared with mock-injected siblings, n = 3, 30 embryos per sample, p d 0.05.

(L) qPCR assay of ifng1-2 and crfb17 expression at 36 hpf in tnnt2 MO-injected embryos compared with mock-injected siblings, n = 3, 30 embryos per sample, p d 0.05.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP la2Tg; s896Tg control Long-pec posterior cardinal vein hematopoietic stem cell IHC
la2Tg; s896Tg + MO1-tnnt2a standard conditions Long-pec posterior cardinal vein hematopoietic stem cell IHC
la2Tg; s896Tg; s994Tg + MO1-tnnt2a heat shock Long-pec posterior cardinal vein hematopoietic stem cell IHC
ifng1 WT control Prim-25 axial vasculature ISH
WT + MO1-tnnt2a standard conditions Prim-25 axial vasculature ISH
ifngr1 WT control Prim-25 axial vasculature ISH
WT + MO1-ifng1 standard conditions Prim-25 axial vasculature ISH
mCherry la2Tg; s896Tg control Long-pec vasculature IHC
Long-pec ventral wall of dorsal aorta hematopoietic stem cell IHC
la2Tg; s896Tg + MO1-tnnt2a standard conditions Long-pec vasculature IHC
Long-pec ventral wall of dorsal aorta hematopoietic stem cell IHC
la2Tg; s896Tg; s994Tg + MO1-tnnt2a heat shock Long-pec vasculature IHC
Long-pec ventral wall of dorsal aorta hematopoietic stem cell IHC
runx1 WT control Prim-5 angioblastic mesenchymal cell ISH
WT + MO1-tnnt2a standard conditions Prim-5 angioblastic mesenchymal cell ISH
s994Tg + MO1-tnnt2a heat shock Prim-5 angioblastic mesenchymal cell ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
la2Tg; s896Tg + MO1-tnnt2a standard conditions Long-pec ventral wall of dorsal aorta has fewer parts of type hematopoietic stem cell, 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