ZFIN ID: ZDB-FIG-200401-10
Kobayashi et al., 2019 - Dual role of Jam3b in early hematopoietic and vascular development. Development (Cambridge, England)   147(1) Full text @ Development
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibodies:
Fish:
Conditions:
Anatomical Terms:
Stage Range: 10-13 somites to Prim-5
PHENOTYPE:
Fish:
Conditions:
Observed In:
Stage Range: 10-13 somites to Prim-5

Fig. 5 Jam3b suppresses Rap1a-Erk activity to promote hemato-vascular development. (A) Confocal imaging of FRET signals under the kdrl:mCherry background. WT and jam3bsa37 embryos injected with Raichu-Rap1 mRNA are shown. Insets show a high magnification view of the dotted boxed area. Arrowheads indicate the kdrl:mCherry (+) vascular cord. Data are representative of eight embryos for each genotype. (B) Western blotting analysis (left) of phosphorylated Erk1/2 (pErk1/2), total Erk1/2 and Tubulin at 23 hpf in WT embryos treated with DMSO and jam3bsa37 embryos treated with DMSO, SL-327 or PD98059. The graph (right) denotes the ratio of pErk1/2 to total Erk1/2 in each embryo. Error bars represent s.d. (n=3 each). (C) Immunostaining of pErk1/2 in WT or jam3bsa37 embryos at 23 hpf (left). The graph (right) denotes the mean intensity of pErk1/2 in WT and jam3bsa37 embryos. Error bars represent s.d. (n=8 each). (D) Immunostaining of pErk1/2 in WT or jam3bsa37 embryos uninjected or co-injected with rap1aa and rap1ab MO (rap1aa/b MO). PLPM cells labeled with fli1a:GFP at 15 hpf are shown. White arrowheads indicate pErk1/2 (+) fli1a:GFP (+) cells. (E) Expression of drl, npas4l, lrrc15 and fli1a in jam3bsa37 embryos treated with DMSO (left) or PD98059 (right). (F) Expression of runx1 in the DA in WT embryos treated with DMSO (left) or in jam3bsa37 embryos treated with DMSO, or PD98059, uninjected or injected with lrrc15 mRNA (right). (G) Expression of drl, npas4l and fli1a in jam3bsa37 embryos treated with DMSO (left) or SU5402 (right). Percent distribution of runx1 expression in F is shown in Fig. S8B. Black arrowheads in E-G indicate the expression domain of each gene. *P<0.05, **P<0.01, ***P<0.001 (unpaired two-tailed Student's t-test). Numbers in bottom right of panels indicate the number of embryos showing the displayed expression pattern over the total number of analyzed embryos. Scale bars: 100 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
drl jam3bsa37/sa37 control 10-13 somites posterior lateral plate mesoderm ISH
jam3bsa37/sa37 chemical treatment: 2-(2-amino-3-methoxyphenyl)chromen-4-one 10-13 somites posterior lateral plate mesoderm ISH
EGFP jam3bsa37; y1Tg standard conditions 10-13 somites endothelial cell IFL
y1Tg standard conditions 10-13 somites endothelial cell IFL
fli1 jam3bsa37/sa37 control 14-19 somites vascular cord ISH
26+ somites dorsal aorta ISH
jam3bsa37/sa37 chemical treatment: 2-(2-amino-3-methoxyphenyl)chromen-4-one 14-19 somites vascular cord ISH
jam3bsa37/sa37 chemical treatment: SU5402 26+ somites dorsal aorta ISH
lrrc15 jam3bsa37/sa37 control 14-19 somites vascular cord ISH
jam3bsa37/sa37 chemical treatment: 2-(2-amino-3-methoxyphenyl)chromen-4-one 14-19 somites vascular cord ISH
npas4l jam3bsa37/sa37 control 10-13 somites posterior lateral plate mesoderm ISH
jam3bsa37/sa37 chemical treatment: 2-(2-amino-3-methoxyphenyl)chromen-4-one 10-13 somites posterior lateral plate mesoderm ISH
runx1 WT control Prim-5 dorsal aorta ISH
jam3bsa37/sa37 control Prim-5 dorsal aorta ISH
jam3bsa37/sa37 chemical treatment: 2-(2-amino-3-methoxyphenyl)chromen-4-one Prim-5 dorsal aorta ISH
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab7-mapk IHC jam3bsa37; y1Tg standard conditions 10-13 somites posterior lateral plate mesoderm
IHC 10-13 somites somite
IHC y1Tg standard conditions 10-13 somites somite
WB WT control 26+ somites whole organism
WB jam3bsa37/sa37 control 26+ somites whole organism
WB jam3bsa37/sa37 chemical treatment: 2-(2-amino-3-methoxyphenyl)chromen-4-one 26+ somites whole organism
WB jam3bsa37/sa37 chemical treatment: SL-327 26+ somites whole organism
Ab13-mapk WB WT control 26+ somites whole organism
WB jam3bsa37/sa37 control 26+ somites whole organism
WB jam3bsa37/sa37 chemical treatment: 2-(2-amino-3-methoxyphenyl)chromen-4-one 26+ somites whole organism
WB jam3bsa37/sa37 chemical treatment: SL-327 26+ somites whole organism
Phenotype Details
Fish Conditions Stage Phenotype
jam3bsa37/sa37 control 26+ somites whole organism ab7-mapk labeling increased amount, abnormal
26+ somites whole organism ab13-mapk labeling increased amount, abnormal
Prim-5 dorsal aorta runx1 expression decreased distribution, abnormal
jam3bsa37/sa37 chemical treatment: 2-(2-amino-3-methoxyphenyl)chromen-4-one 10-13 somites posterior lateral plate mesoderm drl expression increased amount, abnormal
10-13 somites posterior lateral plate mesoderm npas4l expression increased amount, abnormal
14-19 somites vascular cord fli1 expression increased amount, abnormal
14-19 somites vascular cord lrrc15 expression increased amount, abnormal
26+ somites whole organism ab7-mapk labeling increased amount, abnormal
26+ somites whole organism ab13-mapk labeling increased amount, abnormal
jam3bsa37/sa37 chemical treatment: SL-327 26+ somites whole organism ab13-mapk labeling increased amount, abnormal
26+ somites whole organism ab7-mapk labeling increased amount, abnormal
jam3bsa37; y1Tg standard conditions 10-13 somites posterior lateral plate mesoderm ab7-mapk labeling spatial pattern, 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