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ZFIN ID: ZDB-FIG-140801-2
Coxam et al., 2014 - Pkd1 Regulates Lymphatic Vascular Morphogenesis during Development. Cell Reports   7:623-33 Full text @ Cell Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: Prim-5 to Days 7-13
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Day 5

Fig. 2

lyc1 Is a pkd1a Mutant

(A) Overview of positional cloning of lyc1. Individual recombinant embryos (labeled in red left [from 776 embryos analyzed], labeled in blue right [from 314 embryos analyzed]) identify flanking polymorphic markers and limit the critical interval to a region containing partial sequences for pkd1a and tsc2.

(B) Sequence chromatograms showing the wild-type (upper) and pkd1a mutant (R3607X, lower) sequences.

(C) Overall morphology of 5 dpf WT, lyc1, MO-pkd1b, lyc1/MO-pkd1b, and MO-pkd2 embryos. The injection of MO-pkd1b into lyc1 mutants recapitulates the published MO-pkd1a/1b double loss-of-function phenotype ( Mangos et al., 2010).

(D) Expression pattern of pkd1a by in situ hybridization in the trunk of wild-type zebrafish at 24 hpf and 32 hpf.

(E) Quantitative RT-PCR for markers enriched in venous endothelial cells (VECs); kdrl, cdh5, LECs; prox1a, nfatc1, nrp2a; and both flt4 and lyve1 demonstrated the purity of FACS-isolated populations at 5 dpf.

(F) Quantitative RT-PCR for pkd1a, pkd1b, and pkd2 transcripts in 3, 5, and 7 dpf VEC and LEC populations.

(G–I) The vasculature of 5 dpf WT, lyc1/MO-pkd1b and MO-pkd2 embryos (5 and 7.5 ng MO, respectively); asterisk indicates absence of thoracic duct in Tg(fli1a:EGFPy1; kdrl:egfps843) embryos.

Error bars indicate SEM. See also Figures S3 and S4.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
cdh5 nz101Tg; s843Tg standard conditions Day 5 lymph vessel endothelium endothelial cell RTPCR
Day 5 vein endothelial cell RTPCR
EGFP hu5333Tg; y1Tg standard conditions Day 5 thoracic duct IFL
Day 5 trunk vasculature IFL
hu5333Tg; y1Tg + MO4-pkd2 standard conditions Day 5 trunk vasculature IFL
pkd1ahu5855/hu5855; hu5333Tg; y1Tg + MO1-pkd1b standard conditions Day 5 trunk vasculature IFL
flt4 nz101Tg; s843Tg standard conditions Day 5 lymph vessel endothelium endothelial cell RTPCR
Day 5 vein endothelial cell RTPCR
kdrl nz101Tg; s843Tg standard conditions Day 5 lymph vessel endothelium endothelial cell RTPCR
Day 5 vein endothelial cell RTPCR
lyve1b nz101Tg; s843Tg standard conditions Day 5 lymph vessel endothelium endothelial cell RTPCR
Day 5 vein endothelial cell RTPCR
nfatc1 nz101Tg; s843Tg standard conditions Day 5 lymph vessel endothelium endothelial cell RTPCR
Day 5 vein endothelial cell RTPCR
nrp2a nz101Tg; s843Tg standard conditions Day 5 lymph vessel endothelium endothelial cell RTPCR
Day 5 vein endothelial cell RTPCR
pkd1a WT standard conditions Prim-5 whole organism ISH
Prim-15 trunk ISH
nz101Tg; s843Tg standard conditions Protruding-mouth lymph vessel endothelium endothelial cell RTPCR
Protruding-mouth vein endothelial cell RTPCR
Day 5 lymph vessel endothelium endothelial cell RTPCR
Day 5 vein endothelial cell RTPCR
Days 7-13 lymph vessel endothelium endothelial cell RTPCR
Days 7-13 vein endothelial cell RTPCR
pkd1b nz101Tg; s843Tg standard conditions Protruding-mouth lymph vessel endothelium endothelial cell RTPCR
Protruding-mouth vein endothelial cell RTPCR
Day 5 lymph vessel endothelium endothelial cell RTPCR
Day 5 vein endothelial cell RTPCR
Days 7-13 lymph vessel endothelium endothelial cell RTPCR
Days 7-13 vein endothelial cell RTPCR
pkd2 nz101Tg; s843Tg standard conditions Protruding-mouth lymph vessel endothelium endothelial cell RTPCR
Protruding-mouth vein endothelial cell RTPCR
Day 5 lymph vessel endothelium endothelial cell RTPCR
Day 5 vein endothelial cell RTPCR
Days 7-13 lymph vessel endothelium endothelial cell RTPCR
Days 7-13 vein endothelial cell RTPCR
prox1a nz101Tg; s843Tg standard conditions Day 5 vein endothelial cell RTPCR
RFP hu5333Tg; y1Tg standard conditions Day 5 trunk vasculature IFL
hu5333Tg; y1Tg + MO4-pkd2 standard conditions Day 5 trunk vasculature IFL
pkd1ahu5855/hu5855; hu5333Tg; y1Tg + MO1-pkd1b standard conditions Day 5 trunk vasculature IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO4-pkd2 standard conditions Day 5 whole organism curved dorsal, abnormal
hu5333Tg; y1Tg + MO4-pkd2 standard conditions Day 5 thoracic duct absent, abnormal
Day 5 thoracic duct malformed, abnormal
pkd1ahu5855/hu5855 + MO1-pkd1b standard conditions Day 5 whole organism curved dorsal, abnormal
pkd1ahu5855/hu5855; hu5333Tg; y1Tg + MO1-pkd1b standard conditions Day 5 thoracic duct absent, abnormal
Day 5 thoracic duct malformed, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Cell Reports for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Cell Rep.