ZFIN ID: ZDB-FIG-130228-28
Cooney et al., 2013 - Teleost growth factor independence (gfi) genes differentially regulate successive waves of hematopoiesis. Developmental Biology   373(2):431-441 Full text @ Dev. Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: Prim-25 to Day 4
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Day 4

Fig. 5 Loss ofgfi1b silences definitive HSC. (A) Loss of gfi1b silences runx-1, c-myb and ikaros expressing HSC in the AGM at 36 hpf relative to matched controls (a–f). Loss of gfi1b also reduces expression of GFP+cd41 cells in Tg(cd41:eGFP) embryos at 96 hpf (d–f). (B) FACS of Tg(cd41:eGFP) embryos at 96 hpf injected with MO targeting either gfi1aa or gfi1b reveals a marked reduction in the GFPlo and GFPhi populations in gfi1b morphants (mean±SE, t test, p<0.05, n=3). qRT-PCR of gfi morphants shows significantly reduced expression of itga2b (cd41) in gfi1b morphants at 96 hpf (mean±SE, t test, *p<0.05, n=3). (C) Loss of gfi1b silences rag-1 expressing thymic lymphocytes, while loss of gfi1aa has no effect on lymphopoiesis in the thymic anlage (a–c). (d–f) Embryos were stained with Alcian Blue to delineate the morphologic architecture of the jaw cartilages. gfi1aa morphants have normal jaw cartilage development compared with wild type controls (d, e), while gfi1b morphants show dysplastic development of the jaw cartilage (f).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
ikzf1 WT control Prim-25 ventral wall of dorsal aorta ISH
WT + MO1-gfi1aa standard conditions Prim-25 ventral wall of dorsal aorta ISH
WT + MO1-gfi1b standard conditions Prim-25 ventral wall of dorsal aorta ISH
itga2b la2Tg control Day 4 whole organism RTPCR
la2Tg + MO1-gfi1aa standard conditions Day 4 whole organism RTPCR
la2Tg + MO1-gfi1b standard conditions Day 4 whole organism RTPCR
myb WT control Prim-25 ventral wall of dorsal aorta ISH
WT + MO1-gfi1aa standard conditions Prim-25 ventral wall of dorsal aorta ISH
WT + MO1-gfi1b standard conditions Prim-25 ventral wall of dorsal aorta ISH
rag1 WT control Day 4 thymus ISH
WT + MO1-gfi1aa standard conditions Day 4 thymus ISH
WT + MO1-gfi1b standard conditions Day 4 thymus ISH
runx1 WT control Prim-25 ventral wall of dorsal aorta ISH
WT + MO1-gfi1aa standard conditions Prim-25 ventral wall of dorsal aorta ISH
WT + MO1-gfi1b standard conditions Prim-25 ventral wall of dorsal aorta ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
la2Tg + MO1-gfi1aa standard conditions Day 4 hematopoietic stem cell normal amount, normal
Day 4 mandibular arch skeleton morphology, normal
Day 4 thrombocyte increased amount, abnormal
la2Tg + MO1-gfi1b standard conditions Day 4 definitive hemopoiesis disrupted, abnormal
Day 4 hematopoietic stem cell decreased amount, abnormal
Day 4 mandibular arch skeleton dysplastic, abnormal
Day 4 thrombocyte decreased amount, abnormal
Acknowledgments:
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Reprinted from Developmental Biology, 373(2), Cooney, J.D., Hildick-Smith, G.J., Shafizadeh, E., McBride, P.F., Carroll, K.J., Anderson, H., Shaw, G.C., Tamplin, O.J., Branco, D.S., Dalton, A.J., Shah, D.I., Wong, C., Gallagher, P.G., Zon, L.I., North, T.E., and Paw, B.H., Teleost growth factor independence (gfi) genes differentially regulate successive waves of hematopoiesis, 431-441, Copyright (2013) with permission from Elsevier. Full text @ Dev. Biol.