ZFIN ID: ZDB-FIG-151130-5
Kiesow et al., 2015 - Junb controls lymphatic vascular development in zebrafish via miR-182. Scientific Reports   5:15007 Full text @ Sci. Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage: Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Long-pec to Protruding-mouth

Fig. 3 Junb knockout in zebrafish phenocopies the MO-mediated Junb knockdown PL phenotype.

(a) Left panel: Bright field images of representative embryos at 72 hpf injected with the indicated CRISPR gRNA: co-CRISPR (400 pg), junba-CRISPR (250 pg), junbb-CRISPR (250 pg), junba-CRISPR + junbb-CRISPR (200 pg + 200 pg). Middle panel: Confocal images of trunk vasculature in Tg(fli1:EGFP)y1 embryonic region marked by rectangle on the left. Right panel, enlarged view of trunk region marked by the yellow rectangle in the middle panel. DA, dorsal aorta; PCV, posterior cardinal vein; ISVs, intersegmental vessels; PL marked with yellow arrows or if absent with asterisks. Panels are lateral view; dorsal is up, anterior to the left. Scale bars represent 500 µm (black) and 50 µm (white). (b) Percentage of PL defects in embryos (at 48 and 72 hpf) injected with CRISPR gRNAs as described in (a) against either junba (n = 68), junbb (n = 94) or in combination against both (n = 150) versus embryos injected with control CRISPR at corresponding doses of 250 pg (n = 99) or 400 pg (co-CRISPR, n = 105). Sum of 16 hemisegments per embryo was categorized into groups with no, partial or complete absence of PLs. ***P < 0.001, Mann-Whitney U-test. (c) Quantification of arterial and venous ISVs (aISV, vISV) given as % aISV and vISV in Tg(fli1:EGFP)y1 zebrafish embryos at 72 hpf injected with indicated CRISPR gRNAs. n = 40 embryo/group, mean ± SD. **P < 0.01, ns, not significant, Student’s t-test.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP y1Tg control Protruding-mouth dorsal aorta IFL
Protruding-mouth dorsal longitudinal anastomotic vessel IFL
Protruding-mouth intersegmental vessel IFL
Protruding-mouth posterior cardinal vein IFL
Protruding-mouth vascular lymphangioblast IFL
y1Tg + CRISPR1-junba control Protruding-mouth dorsal aorta IFL
Protruding-mouth dorsal longitudinal anastomotic vessel IFL
Protruding-mouth intersegmental vessel IFL
Protruding-mouth posterior cardinal vein IFL
y1Tg + CRISPR1-junba + CRISPR1-junbb control Protruding-mouth dorsal aorta IFL
Protruding-mouth dorsal longitudinal anastomotic vessel IFL
Protruding-mouth intersegmental vessel IFL
Protruding-mouth posterior cardinal vein IFL
y1Tg + CRISPR1-junbb control Protruding-mouth dorsal aorta IFL
Protruding-mouth dorsal longitudinal anastomotic vessel IFL
Protruding-mouth intersegmental vessel IFL
Protruding-mouth posterior cardinal vein IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
y1Tg + CRISPR1-junba control Long-pec lymph vessel development delayed, abnormal
Long-pec vascular lymphangioblast absent, abnormal
Protruding-mouth lymph vessel development delayed, abnormal
Protruding-mouth vascular lymphangioblast absent, abnormal
y1Tg + CRISPR1-junba + CRISPR1-junbb control Long-pec lymph vessel development delayed, abnormal
Long-pec vascular lymphangioblast absent, abnormal
Protruding-mouth lymph vessel development delayed, abnormal
Protruding-mouth vascular lymphangioblast absent, abnormal
y1Tg + CRISPR1-junbb control Long-pec lymph vessel development delayed, abnormal
Long-pec vascular lymphangioblast absent, abnormal
Protruding-mouth lymph vessel development delayed, abnormal
Protruding-mouth vascular lymphangioblast absent, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Scientific Reports for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Sci. Rep.