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ZFIN ID: ZDB-FIG-171207-29
Lu et al., 2015 - Direct regulation of p53 by miR-142a-3p mediates the survival of hematopoietic stem and progenitor cells in zebrafish. Cell discovery   1:15027 Full text @ Cell Discov
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Anatomical Terms:
Stage Range: Prim-5 to Day 4
PHENOTYPE:
Fish:
Observed In:
Stage Range: Prim-5 to Adult

Fig. 2

MiR-142a-3p is essential for HSPC emergence. (a) 142T−/− embryos showed decreased expression of HSPC marker runx1 at 26 and 36 hpf by WISH. (b) 142T−/− embryos displayed decreased expression of HSC marker runx1 at 26 and 36 hpf using qPCR. The expression of runx1 was normalized to β-actin (mean±s.d., n=3, **P<0.05). (c) Runx1 was decreased in 142T−/− embryos at 26 and 36 hpf by western blot analysis (left panel) and the quantification (right panel). The protein level of Runx1 was normalized to β-Actin. (d) The number of EGFP-labeled HSPCs (red arrowheads) in Tg (cmyb:EGFP) embryos was decreased in the AGM region in 142T−/− embryos at 36 hpf (mean±s.d., n=5, *P<0.05). White arrowheads indicate cmyb-positive cells in the pronephric duct. cmyb-labeled pronephric duct was counted as an internal control. (e) 142T−/− embryos displayed reduction of HSPC marker cmyb from 2 to 3.5 dpf by WISH. (f) Differentiated hematopoietic lineages were decreased in 142T−/− embryos including erythrocytes (gata1), neutrophils (lyz) and lymphocytes (ikaros and rag1) at 4 dpf by WISH. Red arrowheads mark hematopoietic cells in the caudal hematopoietic tissue (CHT), whereas red circles denote the thymus. (g) Hematoxylin and eosin (HE) staining of the kidney showed that decrease in cell population and the volume of the head kidney greatly in 142−/− adult fish of 12 weeks.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP zf169Tg standard conditions Prim-25 hematopoietic multipotent progenitor cell IHC
zf169Tg ; mir142aioz13/ioz13 standard conditions Prim-25 hematopoietic multipotent progenitor cell IHC
gata1a WT standard conditions Day 4 caudal hematopoietic tissue hematopoietic multipotent progenitor cell ISH
Day 4 erythroid lineage cell ISH
mir142aioz13/ioz13 standard conditions Day 4 caudal hematopoietic tissue hematopoietic multipotent progenitor cell ISH
Day 4 erythroid lineage cell ISH
ikzf1 WT standard conditions Day 4 lymphocyte ISH
Day 4 thymus ISH
mir142aioz13/ioz13 standard conditions Day 4 lymphocyte ISH
Day 4 thymus ISH
lyz WT standard conditions Day 4 caudal hematopoietic tissue hematopoietic multipotent progenitor cell ISH
Day 4 neutrophil ISH
mir142aioz13/ioz13 standard conditions Day 4 caudal hematopoietic tissue hematopoietic multipotent progenitor cell ISH
Day 4 neutrophil ISH
myb WT standard conditions Long-pec hematopoietic multipotent progenitor cell ISH
Protruding-mouth hematopoietic multipotent progenitor cell ISH
mir142aioz13/ioz13 standard conditions Long-pec hematopoietic multipotent progenitor cell ISH
Protruding-mouth hematopoietic multipotent progenitor cell ISH
rag1 WT standard conditions Day 4 lymphocyte ISH
Day 4 thymus ISH
mir142aioz13/ioz13 standard conditions Day 4 lymphocyte ISH
Day 4 thymus ISH
runx1 WT standard conditions Prim-5 hematopoietic multipotent progenitor cell ISH
Prim-25 hematopoietic multipotent progenitor cell ISH
mir142aioz13/ioz13 standard conditions Prim-5 hematopoietic multipotent progenitor cell ISH
Prim-5 trunk RTPCR
Prim-25 hematopoietic multipotent progenitor cell ISH
Prim-25 trunk RTPCR
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-runx1 WB mir142aioz13/ioz13 standard conditions Prim-5 trunk
WB Prim-25 trunk
Phenotype Details
Fish Conditions Stage Phenotype
mir142aioz13/ioz13 standard conditions Prim-5 hematopoietic multipotent progenitor cell runx1 expression decreased amount, abnormal
Prim-5 hemopoiesis decreased occurrence, abnormal
Prim-5 trunk runx1 expression decreased amount, abnormal
Prim-5 trunk ab1-runx1 labeling decreased amount, abnormal
Prim-25 hematopoietic multipotent progenitor cell runx1 expression decreased amount, abnormal
Prim-25 hemopoiesis decreased occurrence, abnormal
Prim-25 trunk runx1 expression decreased amount, abnormal
Prim-25 trunk ab1-runx1 labeling decreased amount, abnormal
Long-pec hematopoietic multipotent progenitor cell myb expression decreased amount, abnormal
Long-pec hemopoiesis decreased occurrence, abnormal
Protruding-mouth hematopoietic multipotent progenitor cell myb expression decreased amount, abnormal
Protruding-mouth hemopoiesis decreased occurrence, abnormal
Day 4 caudal hematopoietic tissue hematopoietic multipotent progenitor cell lyz expression decreased amount, abnormal
Day 4 caudal hematopoietic tissue hematopoietic multipotent progenitor cell gata1a expression decreased amount, abnormal
Day 4 erythroid lineage cell gata1a expression decreased amount, abnormal
Day 4 hemopoiesis decreased occurrence, abnormal
Day 4 lymphocyte rag1 expression decreased amount, abnormal
Day 4 lymphocyte ikzf1 expression decreased amount, abnormal
Day 4 neutrophil lyz expression decreased amount, abnormal
Day 4 thymus rag1 expression decreased amount, abnormal
Day 4 thymus ikzf1 expression decreased amount, abnormal
Adult head kidney decreased volume, abnormal
zf169Tg ; mir142aioz13/ioz13 standard conditions Prim-25 ventral wall of dorsal aorta hematopoietic multipotent progenitor cell decreased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Cell discovery for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Cell Discov