ZFIN ID: ZDB-FIG-130412-5
Lu et al., 2013 - EpCAM Is an Endoderm-Specific Wnt Derepressor that Licenses Hepatic Development. Developmental Cell   24(5):543-553 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Term:
Stage: Prim-5
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage: Day 4

Fig. 4 EpCAM Directly Binds to Kremen1 and Disrupts the Kremen1-Dkk2 Interaction(A) EpCAM-Myc and Kremen1-HA mutually coimmunoprecipitated from HEK293T cells, negatively controlled by Wnt2bb-HA and Lrp6-HA. IP, immunoprecipitation; IB, immunoblotting.(B) Kremen1-HA expressed in HEK293T cells is specifically pulled down by a GST-EpCAM fusion protein, but not by GST fused to an irrelevant nuclear protein Geminin (GST-Gem). Bmpr1ba-HA and Wnt2bb-HA expressed in HEK293T cells are applied as negative controls. Quantities of GST-Gem and GST-EpCAM recombinant proteins used in the pull-down assays were exhibited.(C) Controlled by preimmune serum, endogenous EpCAM in the embryonic lysate at 24 hpf coimmunoprecipitates with antibodies against Kremen1.(D–F) Expressions of dkk1a (D, 24/25), dkk1b (E, 26/29), and dkk2 (F, 12/15) at 24 hpf. Note that only dkk2 is present in the bilateral LPM domains adjacent to the hepatic endoderm (arrowheads).(G–L) Under the Tg(lfabp:GFP) background at 102 hpf, defects in liver development caused by dkk2 mRNA (H, 44/73) or kremen1 mRNA (J, 47/87) are rescued by epcam mRNA (I, 53/64) and (K, 99/135). Reduced liver size in hi2151 mutants is rescued by dkk2MO (L, 50/76).(M) In HEK293T cells, along with increases in the amount of epcam-Myc plasmid used for transfection (μg), the levels of coimmunoprecipitated Dkk2-Flag with Kremen1-HA get reduced.(N) In HEK293T cells, along with increases in the amount of dkk2-Flag plasmid used for transfection (μg), the levels of coimmunoprecipitated EpCAM-Myc with Kremen1-HA get reduced.See also Figures S1, S5, and Table S1.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
dkk1a WT standard conditions Prim-5 Not Detected lateral plate mesoderm ISH
dkk1b WT standard conditions Prim-5 Not Detected lateral plate mesoderm ISH
dkk2 WT standard conditions Prim-5 lateral plate mesoderm ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
epcamhi2151Tg; as3Tg + MO1-dkk2 standard conditions Day 4 liver size, normal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Cell for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Developmental Cell, 24(5), Lu, H., Ma, J., Yang, Y., Shi, W., and Luo, L., EpCAM Is an Endoderm-Specific Wnt Derepressor that Licenses Hepatic Development, 543-553, Copyright (2013) with permission from Elsevier. Full text @ Dev. Cell