ZFIN ID: ZDB-FIG-170824-19
Chiang et al., 2017 - SoxF factors induce Notch1 expression via direct transcriptional regulation during early arterial development. Development (Cambridge, England)   144(14):2629-2639 Full text @ Development
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Terms:
Stage: Prim-5
PHENOTYPE:
Fish:
Condition:
Knockdown Reagent:
Observed In:
Stage Range: Prim-5 to Protruding-mouth

Fig. 8

Loss of endogenous notch1b-15 compromises artery formation and reduces the endogenous notch1b transcript level. (A) The deleted region (dashed line) of notch1b-15 mutant allele uq1mf, which includes both the zfSOX-a and zfSOX-b sites (yellow). (B) F2 notch1b-15uq1mf/uq1mf has reduced notch1b expression in dorsal aorta and intersomitic vessels (white arrows) as compared with sibling WT and heterozygotes (black arrows) at 26-28 hpf. The number of embryos showing the illustrated phenotype among the total examined is indicated. (C) Quantitative PCR on FACS-sorted endothelial populations at 24-28 hpf, showing that F3 notch1b-15uq1mf/uq1mf fish have lower notch1b expression than WT fish. Expression is relative to kdrl and flt1. Mean±s.e.m. n=6 (uq1mf/uq1mf) and n=8 (WT) independent sorts, where each sort was pooled from 60-100 larvae;. *P<0.05, ***P<0.001 (t-test). (D) The treatment regime conducted to characterise the vascular phenotype of the uq1mf/+ cross. In treatment 1 (T1, red), notch1b MO was injected at the 1-2 cell stage. The developing vasculature of each embryo was then analysed blindly at 3 dpf. After scoring, genotypes were assigned to each larva. In treatment 2 (T2, blue), larvae from the uq1mf/+ cross were treated with or without DAPT (5 µM) from 15-16 hpf until 3 dpf. Vessels of these treated larvae were blindly scored prior to genotyping, as reported for T1. (E) At 3 dpf, notch1b-15uq1mf/uq1mf notch1b morphants frequently showed ectopic sprouting in between the intersomitic vessels (asterisks) as compared with sibling WT notch1b morphants. Mutants also show loss of arterial connections (red arrowheads) between the dorsal longitudinal anastomotic vessel (DLAV) and dorsal aorta (DA), as indicated by the loss of YFP expression in the tg(flt1:YFP) background. (F) (Top) Quantification of hypersprouting ISV number in individual notch1b morphants labelled by tg(flt1:YFP) at 3 dpf. Mean±s.e.m. Sibling WT (+/+), n=20; heterozygote (uq1mf/+), n=32; homozygous mutant (uq1mf/uq1mf), n=21. (Bottom) Quantification of YFP-positive intersomitic vessels that connect between DLAV and DA in individual notch1b morphants labelled by tg(flt1:YFP) at 3 dpf. Mean±s.e.m. Sibling WT, n=20; heterozygote, n=32; homozygous mutant, n=21. **P<0.005 (Mann–Whitney U-test). (G) Quantification of YFP-positive intersomitic vessels that connect between DLAV and DA in individual embryos treated with 5 μM DAPT at 3 dpf. Vessels are labelled by tg(flt1:YFP). Mean±s.e.m. Sibling WT, n=9; heterozygote, n=25; homozygous mutant, n=14. *P<0.05 (Mann–Whitney U-test).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
dll4 hu4624Tg; nz101Tg standard conditions Prim-5 endothelial cell RTPCR
notch1bzf2046/zf2046 ; nz101Tg ; hu4624Tg standard conditions Prim-5 endothelial cell RTPCR
flt1 hu4624Tg; nz101Tg standard conditions Prim-5 endothelial cell RTPCR
notch1bzf2046/zf2046 ; nz101Tg ; hu4624Tg standard conditions Prim-5 endothelial cell RTPCR
her6 hu4624Tg; nz101Tg standard conditions Prim-5 endothelial cell RTPCR
notch1bzf2046/zf2046 ; nz101Tg ; hu4624Tg standard conditions Prim-5 endothelial cell RTPCR
hey1 hu4624Tg; nz101Tg standard conditions Prim-5 endothelial cell RTPCR
notch1bzf2046/zf2046 ; nz101Tg ; hu4624Tg standard conditions Prim-5 endothelial cell RTPCR
hey2 hu4624Tg; nz101Tg standard conditions Prim-5 endothelial cell RTPCR
notch1bzf2046/zf2046 ; nz101Tg ; hu4624Tg standard conditions Prim-5 endothelial cell RTPCR
kdrl hu4624Tg; nz101Tg standard conditions Prim-5 endothelial cell RTPCR
notch1bzf2046/zf2046 ; nz101Tg ; hu4624Tg standard conditions Prim-5 endothelial cell RTPCR
notch1a hu4624Tg; nz101Tg standard conditions Prim-5 endothelial cell RTPCR
notch1bzf2046/zf2046 ; nz101Tg ; hu4624Tg standard conditions Prim-5 endothelial cell RTPCR
notch1b hu4624Tg; nz101Tg standard conditions Prim-5 dorsal aorta ISH
Prim-5 endothelial cell RTPCR
Prim-5 intersegmental vessel ISH
Prim-5 neural tube ISH
notch1bzf2046/zf2046 ; nz101Tg ; hu4624Tg standard conditions Prim-5 endothelial cell RTPCR
nz101Tg ; hu4624Tg ; notch1bzf2046/zf2046 standard conditions Prim-5 dorsal aorta ISH
Prim-5 intersegmental vessel ISH
Prim-5 neural tube ISH
notch2 hu4624Tg; nz101Tg standard conditions Prim-5 endothelial cell RTPCR
notch1bzf2046/zf2046 ; nz101Tg ; hu4624Tg standard conditions Prim-5 endothelial cell RTPCR
notch3 hu4624Tg; nz101Tg standard conditions Prim-5 endothelial cell RTPCR
notch1bzf2046/zf2046 ; nz101Tg ; hu4624Tg standard conditions Prim-5 endothelial cell RTPCR
sox18 hu4624Tg; nz101Tg standard conditions Prim-5 endothelial cell RTPCR
notch1bzf2046/zf2046 ; nz101Tg ; hu4624Tg standard conditions Prim-5 endothelial cell RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
notch1bzf2046/zf2046 ; nz101Tg ; hu4624Tg standard conditions Prim-5 endothelial cell notch1a expression decreased amount, abnormal
Prim-5 endothelial cell notch1b expression decreased amount, abnormal
nz101Tg ; hu4624Tg ; notch1bzf2046/zf2046 standard conditions Prim-5 dorsal aorta notch1b expression decreased amount, abnormal
Prim-5 intersegmental vessel notch1b expression decreased amount, abnormal
nz101Tg ; hu4624Tg ; notch1bzf2046/zf2046 chemical treatment: DAPT Protruding-mouth intersegmental artery decreased amount, abnormal
nz101Tg ; hu4624Tg ; notch1bzf2046/zf2046 + MO2-notch1b standard conditions Protruding-mouth angiogenic sprout increased amount, abnormal
Protruding-mouth intersegmental artery decreased amount, abnormal
Protruding-mouth sprouting angiogenesis increased process quality, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Development (Cambridge, England) for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Development