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ZFIN ID: ZDB-FIG-180425-9
Singh et al., 2017 - Etv2-miR-130a-Jarid2 cascade regulates vascular patterning during embryogenesis. PLoS One   12:e0189010 Full text @ PLoS One
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage: Long-pec
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Long-pec to Protruding-mouth

Fig. 4

miR-130a regulates endothelial patterning in vivo.

A, B, qPCR analysis of endothelial transcripts, kdr1 and tek at 48hpf using RNA from control and miR-130a morphants. C, Whole-mount in situ hybridization images of control and miR-130a morphants at 48 hpf using cdh5 and kdr1 probes. Note the defective nature of vascular development and reduced expression of these transcripts in miR-130a morphants (arrowheads). D, E, Brightfield microscopic images revealed no major changes in gross morphology of mismatch control and miR-130a morphants. F-I, Lateral fluorescence images of Tg(fli1a:EGFP) zebrafish lines revealed defective vasculature in miR-130a morphants (white arrowheads) at 48 hpf (F, G) and 72 hpf (H, I) time periods. J, K, Quantitative analysis of the number of defective zebrafish embryos with perturbed inter-somitic vessels (ISVs) at 48 hpf and 72 hpf. L, M, ImageJ (L) and FACS (M) analyses of EGFP+ cells revealed significantly reduced EGFP intensity and counts in miR-130a morphants. N, Lateral fluorescence and brightfield images of Tg(fli1a:EGFP) zebrafish lines co-injected with miR-130a morpholinos and LNA modified scrambled oligos and miR-130a mimics at 72hpf. Panel N’ and N” shows the enlarged images of the boxed area in panel N. Note the restoration of the vascular structures following co-injection with miR-130a morpholinos and miR-130a mimics (N’, N”; arrowheads). O, Quantitative analysis of the number of defective zebrafish embryos with perturbed inter-somitic vessels (ISVs) 72 hpf. Error bars indicate SEM (n = 3; *p<0.05; **p < 0.01). Scale bar: 200 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
cdh5 y1Tg standard conditions Long-pec intersegmental vessel ISH
y1Tg + MO1-mir130a-1 standard conditions Long-pec intersegmental vessel ISH
EGFP y1Tg + MO1-mir130a-1 standard conditions Long-pec vasculature IFL
kdrl y1Tg standard conditions Long-pec intersegmental vessel ISH
y1Tg + MO1-mir130a-1 standard conditions Long-pec endothelial cell RTPCR
Long-pec intersegmental vessel ISH
tek y1Tg + MO1-mir130a-1 standard conditions Long-pec endothelial cell RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
y1Tg + MO1-mir130a-1 standard conditions Long-pec endothelial cell tek expression decreased amount, abnormal
Long-pec endothelial cell kdrl expression decreased amount, abnormal
Long-pec intersegmental vessel kdrl expression decreased amount, abnormal
Long-pec intersegmental vessel cdh5 expression decreased amount, abnormal
Long-pec intersegmental vessel incomplete structure, abnormal
Long-pec trunk vasculature morphology, abnormal
Long-pec vasculature EGFP expression decreased amount, abnormal
Long-pec whole organism morphology, normal
Protruding-mouth intersegmental vessel incomplete structure, abnormal
Acknowledgments:
ZFIN wishes to thank the journal PLoS One for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS One