ZFIN ID: ZDB-FIG-170315-9
Zhang et al., 2015 - G protein-coupled receptor 183 facilitates endothelial-to-hematopoietic transition via Notch1 inhibition. Cell Research   25:1093-107 Full text @ Cell Res.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Conditions:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Prim-5 to High-pec
PHENOTYPE:
Fish:
Conditions:
Knockdown Reagent:
Observed In:
Stage Range: Prim-5 to Prim-25

Fig. 3

Downregulation of Notch signaling is required for definitive hematopoiesis. (A) The increase in hey2 expression in gpr183 mutant is reversed by DBZ treatment. Right panel shows the qPCR quantification. (B) The decrease in cmyb expression in gpr183 mutant is reversed by DBZ treatment. In A and B, red arrowheads indicate hey2 or cmyb expression in the AGM region at 33 hpf. (C) Dot-plot analysis of cmyb+ cell numbers in AGM. (D) Clotrimazole treatment increases expression levels of her6 and her9 in flk1+runx1+ cells at 28 hpf. (E) Notch-active endothelial cells increase in gpr183 morphant (n = 15 for control and morpholino treatment). Right panel shows the quantification analysis. (F) EHT from 37 (shown as 00:00:00) to 43 hpf in Tg(runx1:en-GFP/tp1:mCherry) embryo. Asterisks label individual cells. (G, H) cmyb expression in the AGM region of control embryos and embryos in which Notch signaling is inhibited. Tg(hsp70:dn-MAML) embryos expressing a dominant negative (dn) isoform of the murine mastermind-like (MAML) protein fused to GFP under a hsp70 promoter were heat shocked at 20 or 26 hpf; cmyb expression was examined at 36 hpf. GFP embryo serves as a control. Representative images are shown in G and the dot-plot quantification analysis is shown in H. (I, J) cmyb expression in the AGM region of control and Notch signaling-activated embryos. Tg(UAS:NICD) (NICD, control) and Tg(hsp70:GAL4/UAS:NICD) (hs:NICD) embryos were heat shocked at 20 or 26 hpf and cmyb expression was examined at 36 hpf. Representative images are shown in I, and the dot-plot quantification analysis is shown in J. In G and I, red arrowheads mark the staining signals in AGM. Error bar represents SD. *P < 0.05, **P< 0.01, ***P < 0.001.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP jh11Tg; y1Tg control Prim-25 ventral wall of dorsal aorta IFL
jh11Tg; y1Tg + MO1-gpr183a control Prim-25 ventral wall of dorsal aorta IFL
GFP jh11Tg ; ioz1Tg control Prim-25 to High-pec angioblastic mesenchymal cell IFL
Prim-25 to High-pec hematopoietic multipotent progenitor cell IFL
her6 ioz1Tg; s896Tg chemical treatment: clotrimazole Prim-5 angioblastic mesenchymal cell RTPCR
her9 ioz1Tg; s896Tg chemical treatment: clotrimazole Prim-5 angioblastic mesenchymal cell RTPCR
hey2 AB/TU control Prim-15 ventral wall of dorsal aorta ISH
gpr183aioz9/ioz9(AB/TU) control Prim-15 ventral wall of dorsal aorta ISH
Prim-15 whole organism RTPCR
gpr183aioz9/ioz9(AB/TU) chemical treatment: LY-411575 Prim-15 ventral wall of dorsal aorta ISH
Prim-15 whole organism RTPCR
mCherry jh11Tg ; ioz1Tg control Prim-25 to High-pec ventral wall of dorsal aorta IFL
jh11Tg; y1Tg control Prim-25 ventral wall of dorsal aorta IFL
jh11Tg; y1Tg + MO1-gpr183a control Prim-25 ventral wall of dorsal aorta IFL
myb AB/TU control Prim-15 ventral wall of dorsal aorta ISH
WT control Prim-25 ventral wall of dorsal aorta ISH
fb11Tg heat shock Prim-25 ventral wall of dorsal aorta ISH
gpr183aioz9/ioz9(AB/TU) control Prim-15 ventral wall of dorsal aorta ISH
gpr183aioz9/ioz9(AB/TU) chemical treatment: LY-411575 Prim-15 ventral wall of dorsal aorta ISH
kca3Tg heat shock Prim-25 ventral wall of dorsal aorta ISH
kca3Tg; kca4Tg heat shock Prim-25 ventral wall of dorsal aorta ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
fb11Tg heat shock Prim-25 ventral wall of dorsal aorta myb expression decreased amount, abnormal
Prim-25 ventral wall of dorsal aorta myb expression increased amount, abnormal
gpr183aioz9/ioz9(AB/TU) control Prim-15 ventral wall of dorsal aorta hey2 expression increased amount, abnormal
Prim-15 ventral wall of dorsal aorta myb expression decreased amount, abnormal
Prim-15 ventral wall of dorsal aorta Notch signaling pathway increased occurrence, abnormal
Prim-15 whole organism hey2 expression increased amount, abnormal
gpr183aioz9/ioz9(AB/TU) chemical treatment: LY-411575 Prim-15 ventral wall of dorsal aorta hey2 expression amount, ameliorated
Prim-15 ventral wall of dorsal aorta myb expression amount, ameliorated
Prim-15 whole organism hey2 expression amount, ameliorated
ioz1Tg; s896Tg chemical treatment: clotrimazole Prim-5 angioblastic mesenchymal cell her6 expression increased amount, abnormal
Prim-5 angioblastic mesenchymal cell her9 expression increased amount, abnormal
jh11Tg; y1Tg control Prim-25 ventral wall of dorsal aorta mCherry expression increased amount, abnormal
jh11Tg; y1Tg + MO1-gpr183a control Prim-25 ventral wall of dorsal aorta mCherry expression increased amount, abnormal
kca3Tg; kca4Tg heat shock Prim-25 ventral wall of dorsal aorta myb expression decreased amount, abnormal
Prim-25 ventral wall of dorsal aorta myb expression increased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Cell Research for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Cell Res.