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ZFIN ID: ZDB-FIG-161201-35
Zhou et al., 2016 - BLOS2 negatively regulates Notch signaling during neural and hematopoietic stem and progenitor cell development. eLIFE   5 Full text @ Elife
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Terms:
Stage Range: Prim-5 to Day 4
PHENOTYPE:
Fish:
Observed In:
Stage Range: Prim-5 to Day 4

Fig. 5

bloc1s2 deficiency promotes HSPC production in zebrafish.

(A-C) The results of (A) whole mount in situ hybridization (WISH), (B) qRT-PCR and (C) immunoblotting (Figure 5—source data 1) showed that the expression of bloc1s2 at the RNA and protein levels in bloc1s2 mutants was decreased but not abolished compared to that in control embryos at 24 hpf. Each bar represents the mean ± s.e.m of three independent experiments. **p<0.01, ***p<0.001 (Student’s t-test). (D) The Runx1 protein level was increased in bloc1s2 mutants (Figure 5—source data 2). The right panel is the quantitative analysis of the western blotting results. Each bar represents the mean ± s.e.m of three independent experiments. ***p<0.001. (E) The WISH results showed that the expression of runx1 at 24 hpf and 36 hpf, and cmyb expression at 36 hpf were increased in bloc1s2 mutants. Black arrowheads mark expression of runx1 and cmyb in the AGM region. (F) qRT-PCR results from the dissected trunk region showed that expression of cmyb at 36 hpf was significantly increased in bloc1s2 mutants. Each bar represents the mean ± s.e.m of three independent experiments. *p<0.05. (G) We generated the bloc1s2 mutants in a Tg (flk1:mcherry;cmyb:GFP) background. The upper panels show that the number of flk1+cmyb+ cells in outcrossed embryos was increased. White arrowheads mark flk1+cmyb+ cells in the AGM region at 36 hpf. The lower panel is the quantitative analysis of the western blotting results. Each bar represents the mean ± s.e.m of seven embryos. **p<0.01. (H) rag1 expression was increased in the thymus region, but gata1 and pu1 expression was decreased in the caudal hematopoietic tissue (CHT) region in bloc1s2 mutants at 4.5 dpf. Black dashed circles mark rag1 expression in the thymus region. Black arrowheads mark expression of gata1 and pu1 in the CHT region.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
bloc1s2 TU control Prim-5 endothelial cell ISH
Prim-5 neuron ISH
bloc1s2ioz8/ioz8 standard conditions Prim-5 endothelial cell ISH
Prim-5 neuron ISH
Prim-5 whole organism WB
Prim-5 whole organism RTPCR
EGFP bloc1s2ioz8/ioz8; s896Tg; zf169Tg standard conditions Prim-25 ventral wall of dorsal aorta hematopoietic stem cell IFL
s896Tg; zf169Tg standard conditions Prim-25 ventral wall of dorsal aorta hematopoietic stem cell IFL
gata1a TU control Day 4 erythroid lineage cell ISH
bloc1s2ioz8/ioz8 standard conditions Day 4 erythroid lineage cell ISH
mCherry bloc1s2ioz8/ioz8; s896Tg; zf169Tg standard conditions Prim-25 ventral wall of dorsal aorta hematopoietic stem cell IFL
s896Tg; zf169Tg standard conditions Prim-25 ventral wall of dorsal aorta hematopoietic stem cell IFL
myb TU control Prim-25 ventral wall of dorsal aorta hematopoietic stem cell ISH
bloc1s2ioz8/ioz8 standard conditions Prim-25 ventral wall of dorsal aorta hematopoietic stem cell ISH
Prim-25 whole organism RTPCR
rag1 TU control Day 4 thymus lymphoid progenitor cell ISH
bloc1s2ioz8/ioz8 standard conditions Day 4 thymus lymphoid progenitor cell ISH
runx1 TU control Prim-5 ventral wall of dorsal aorta hematopoietic stem cell ISH
Prim-25 ventral wall of dorsal aorta hematopoietic stem cell ISH
bloc1s2ioz8/ioz8 standard conditions Prim-5 ventral wall of dorsal aorta hematopoietic stem cell ISH
Prim-5 whole organism WB
Prim-25 ventral wall of dorsal aorta hematopoietic stem cell ISH
spi1b TU control Day 4 myeloid cell ISH
bloc1s2ioz8/ioz8 standard conditions Day 4 myeloid cell ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
bloc1s2ioz8/ioz8 standard conditions Prim-5 endothelial cell bloc1s2 expression decreased amount, abnormal
bloc1s2ioz8/ioz8 standard conditions Prim-5 neuron bloc1s2 expression decreased amount, abnormal
bloc1s2ioz8/ioz8 standard conditions Prim-5 ventral wall of dorsal aorta hematopoietic stem cell runx1 expression increased amount, abnormal
bloc1s2ioz8/ioz8 standard conditions Prim-5 whole organism bloc1s2 expression decreased amount, abnormal
bloc1s2ioz8/ioz8 standard conditions Prim-5 whole organism bloc1s2 expression decreased amount, abnormal
bloc1s2ioz8/ioz8 standard conditions Prim-5 whole organism runx1 expression increased amount, abnormal
bloc1s2ioz8/ioz8 standard conditions Prim-25 ventral wall of dorsal aorta hematopoietic stem cell runx1 expression increased amount, abnormal
bloc1s2ioz8/ioz8 standard conditions Prim-25 ventral wall of dorsal aorta hematopoietic stem cell myb expression increased amount, abnormal
bloc1s2ioz8/ioz8 standard conditions Prim-25 whole organism myb expression increased amount, abnormal
bloc1s2ioz8/ioz8 standard conditions Day 4 erythroid lineage cell gata1a expression decreased amount, abnormal
bloc1s2ioz8/ioz8 standard conditions Day 4 erythroid lineage cell myeloid progenitor cell differentiation decreased occurrence, abnormal
bloc1s2ioz8/ioz8 standard conditions Day 4 lymphoid progenitor cell differentiation increased occurrence, abnormal
bloc1s2ioz8/ioz8 standard conditions Day 4 myeloid cell spi1b expression decreased amount, abnormal
bloc1s2ioz8/ioz8 standard conditions Day 4 myeloid progenitor cell differentiation decreased occurrence, abnormal
bloc1s2ioz8/ioz8 standard conditions Day 4 thymus lymphoid progenitor cell rag1 expression increased amount, abnormal
bloc1s2ioz8/ioz8; s896Tg; zf169Tg standard conditions Prim-25 ventral wall of dorsal aorta hematopoietic stem cell increased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal eLIFE for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Elife