FIGURE

Fig. 4

ID

ZDB-FIG-130412-5

Publication

Lu et al., 2013 - EpCAM Is an Endoderm-Specific Wnt Derepressor that Licenses Hepatic Development

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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.

Expression Data

Genes:	dkk1a dkk1b dkk2
Fish:	WT
Anatomical Term:	lateral plate mesoderm
Stage:	Prim-5

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	epcam^hi2151Tg; as3Tg + MO1-dkk2
Knockdown Reagent:	MO1-dkk2
Observed In:	liver
Stage:	Day 4

Phenotype Detail

Acknowledgments

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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