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ZFIN ID: ZDB-FIG-190410-12
Meng et al., 2017 - The pro-inflammatory signalling regulator Stat4 promotes vasculogenesis of great vessels derived from endothelial precursors. Nature communications   8:14640 Full text @ Nat. Commun.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Conditions:
Knockdown Reagents:
Anatomical Terms:
Stage: High-pec
PHENOTYPE:
Fish:
Conditions:
Knockdown Reagents:
Observed In:
Stage: High-pec

Fig. 7 stat4 promoted the PAA angioblast programme by suppressing stat1a and hdac3.

(a–k) In situ hybridization analysis of tie1 transcripts in the control (a), stat4 mutants (b), embryos with 50 ng μl−1 pias2 mRNA injection (c), embryos with 50 ng μl−1 socs3a and socs3b mRNAs injection (d), embryos treated with 85 μM lisofylline (e), stat4 mutants with 0.2 μM TSA treatment (f), embryos with 50 ng μl−1 hdac3 mRNA injection (g), stat4 mutants with 4 ng μl−1 hdac3 morpholino (h), stat4 mutants with 4 ng μl−1 stat1b morpholino (i), stat4 mutants with 4 ng μl−1 stat1a morpholino (j) at 44 hpf. Scale bars, 50 μm. (k) Proportional quantification of indicated tie1+ expressing cords number (s4−/−: stat4 mutants), Kruskal–Wallis test with the Dunn’s multiple comparison test, **P<0.01, n.s.: P>0.05, n ≥30 embryos per each group. (l–n) The ChIP analysis with anti-Stat4 antibody for the promoter regions of hdac3 and stat1a in the wild-type control and stat4 mutants at 44 hpf across six experimental replicates (n=30 embryos/replicate in each group, analysis of variance (ANOVA) test with multiple comparison post-hoc test, **P<0.01, ***P<0.001). (m) The binding enrichment regions at the hdac3 and stat1a locus are indicated by primers (P1–P6). (o) Q-PCR results of related genes expression in the control and stat4 morphants at 44 hpf across six experimental replicates (n=10 embryos/replicate in each group. Error bars indicate s.d., unpaired two-tailed Student’s t-test, *P<0.05, **P<0.01. (p) Protein levels of Nkx2.5, Hdac3 and Stat1a in the wild-type control and stat4 mutants detected by western blots at 44 hpf. β-actin is used as the internal control. (q) A network is drawn to illustrate the stat4 pathway in controlling PAA proliferation. Red: activation, Blue: suppression.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
atrip AB/TU standard conditions High-pec whole organism RTPCR
AB/TU + MO1-stat4 standard conditions High-pec whole organism RTPCR
cdk2 AB/TU standard conditions High-pec whole organism RTPCR
AB/TU + MO1-stat4 standard conditions High-pec whole organism RTPCR
cdkn1ca AB/TU standard conditions High-pec whole organism RTPCR
AB/TU + MO1-stat4 standard conditions High-pec whole organism RTPCR
cdkn2a/b AB/TU standard conditions High-pec whole organism RTPCR
AB/TU + MO1-stat4 standard conditions High-pec whole organism RTPCR
hdac3 AB/TU standard conditions High-pec whole organism WB
High-pec whole organism RTPCR
AB/TU + MO1-stat4 standard conditions High-pec whole organism RTPCR
stat4zf2070/zf2070 standard conditions High-pec whole organism WB
nkx2.5 AB/TU standard conditions High-pec whole organism WB
stat4zf2070/zf2070 standard conditions High-pec whole organism WB
stat1a AB/TU standard conditions High-pec whole organism WB
High-pec whole organism RTPCR
AB/TU + MO1-stat4 standard conditions High-pec whole organism RTPCR
stat4zf2070/zf2070 standard conditions High-pec whole organism WB
stat1b AB/TU standard conditions High-pec whole organism RTPCR
AB/TU + MO1-stat4 standard conditions High-pec whole organism RTPCR
tie1 AB/TU standard conditions High-pec aortic arch angioblastic mesenchymal cell ISH
AB/TU chemical treatment by environment: (R)-lisofylline High-pec aortic arch angioblastic mesenchymal cell ISH
stat4zf2070/zf2070 standard conditions High-pec aortic arch angioblastic mesenchymal cell ISH
stat4zf2070/zf2070 chemical treatment by environment: trichostatin A High-pec aortic arch angioblastic mesenchymal cell ISH
stat4zf2070/zf2070 + MO2-stat1a standard conditions High-pec aortic arch angioblastic mesenchymal cell ISH
stat4zf2070/zf2070 + MO2-stat1b standard conditions High-pec aortic arch angioblastic mesenchymal cell ISH
stat4zf2070/zf2070 + MO3-hdac3 standard conditions High-pec aortic arch angioblastic mesenchymal cell ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
AB/TU chemical treatment by environment: (R)-lisofylline High-pec aortic arch 3 angioblast cell differentiation occurrence, normal
High-pec aortic arch 4 angioblast cell differentiation occurrence, normal
High-pec aortic arch 5 angioblast cell differentiation occurrence, normal
High-pec aortic arch 6 apoptotic process occurrence, normal
AB/TU + MO1-stat4 standard conditions High-pec whole organism stat1b expression decreased amount, abnormal
High-pec whole organism cdk2 expression decreased amount, abnormal
High-pec whole organism cdkn1ca expression decreased amount, abnormal
High-pec whole organism cdkn2a/b expression increased amount, abnormal
High-pec whole organism hdac3 expression increased amount, abnormal
High-pec whole organism stat1a expression increased amount, abnormal
High-pec whole organism atrip expression increased amount, abnormal
stat4zf2070/zf2070 standard conditions High-pec aortic arch 3 angioblast cell differentiation decreased occurrence, abnormal
High-pec aortic arch 4 angioblast cell differentiation decreased occurrence, abnormal
High-pec aortic arch 5 angioblast cell differentiation decreased occurrence, abnormal
High-pec aortic arch 6 angioblast cell differentiation decreased occurrence, abnormal
High-pec aortic arch angioblastic mesenchymal cell tie1 expression decreased amount, abnormal
High-pec whole organism stat1a expression increased amount, abnormal
High-pec whole organism hdac3 expression increased amount, abnormal
stat4zf2070/zf2070 chemical treatment by environment: trichostatin A High-pec aortic arch 3 angioblast cell differentiation occurrence, ameliorated
High-pec aortic arch 4 angioblast cell differentiation occurrence, ameliorated
High-pec aortic arch 5 angioblast cell differentiation occurrence, ameliorated
High-pec aortic arch 6 angioblast cell differentiation occurrence, ameliorated
High-pec aortic arch angioblastic mesenchymal cell tie1 expression amount, ameliorated
stat4zf2070/zf2070 + MO2-stat1a standard conditions High-pec aortic arch 3 angioblast cell differentiation occurrence, ameliorated
High-pec aortic arch 4 angioblast cell differentiation occurrence, ameliorated
High-pec aortic arch 5 angioblast cell differentiation occurrence, ameliorated
High-pec aortic arch 6 angioblast cell differentiation occurrence, ameliorated
High-pec aortic arch angioblastic mesenchymal cell tie1 expression amount, ameliorated
stat4zf2070/zf2070 + MO2-stat1b standard conditions High-pec aortic arch 3 angioblast cell differentiation decreased occurrence, abnormal
High-pec aortic arch 4 angioblast cell differentiation decreased occurrence, abnormal
High-pec aortic arch 5 angioblast cell differentiation decreased occurrence, abnormal
High-pec aortic arch 6 angioblast cell differentiation decreased occurrence, abnormal
High-pec aortic arch angioblastic mesenchymal cell tie1 expression decreased amount, abnormal
stat4zf2070/zf2070 + MO3-hdac3 standard conditions High-pec aortic arch 3 angioblast cell differentiation occurrence, ameliorated
High-pec aortic arch 4 angioblast cell differentiation occurrence, ameliorated
High-pec aortic arch 5 angioblast cell differentiation occurrence, ameliorated
High-pec aortic arch 6 angioblast cell differentiation occurrence, ameliorated
High-pec aortic arch angioblastic mesenchymal cell tie1 expression amount, ameliorated
Acknowledgments:
ZFIN wishes to thank the journal Nature communications for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Nat. Commun.