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Fig. 2

ID

ZDB-FIG-150407-2

Publication

Renz et al., 2015 - Regulation of β1 Integrin-Klf2-Mediated Angiogenesis by CCM Proteins

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Fig. 2

Elevated Levels of klf2 mRNA Cause Morphogenetic Defects in Zebrafish ccm Mutants

(A–I) Rescue of cardiac malformations, LDA defects, and SIV defects in zebrafish ccm2^m201 mutants (B, B′, E, and H) by targeting klf2a/b with MOs (C, C′, F, and I). (A–C) Shown are hearts of different genotypes at 48 hrpf or (A′, B′, and C′) details with single confocal plane sections of the atrioventricular canal (AVC) region (arrowheads, white box). Endocardial cushion cells are marked by Tg(kdrl:GFP)^s843 and Alcam staining (asterisks). (D–F) Dorsal views of the LDA marked by Tg(kdrl:GFP)^s843 (inverted images) in different genotypes at 48 hrpf. (G–I) Side view of the SIV marked by Tg(kdrl:GFP)^s843 in different genotypes at 72 hrpf. Vessel branch point defects (arrows) within the SIV are reduced by targeting klf2a/b in ccm2^m201 mutants (H and I).

(J) Endocardial and LDA cell numbers are elevated in ccm2^m201 mutants at 48 hrpf, as well as upon overexpression of klf2a from a heat-shock-inducible promoter.

(K) Quantifications of the number of branch points of the SIV in different genotypes at 72 hrpf.

(L and M) Endothelial overexpression of klf2a in Tg(fli1ep:Gal4FF)^ubs2; Tg(UAS:klf2a)^ig1–3; Tg(kdrl:GFP)^s843 transgenic embryos is sufficient to phenocopy the ccm2^m201 mutant cardiac ballooning phenotype. Endo, endocardium; myo, myocardium; A, atrium; V, ventricle. Scale bars, 25 µm. Statistical data are means and SEM; ns, not significant; p < 0.05; p < 0.01; p < 0.001; p < 0.0001; see Supplemental Experimental Procedures for details about statistical data analysis.

Expression Data

Genes:	alcama GFP
Antibody:	zn-8
Fish:	s843Tg ccm2^m201/m201; s843Tg ccm2^m201/m201; s843Tg + MO1-klf2a + MO2-klf2b
Knockdown Reagents:	MO1-klf2a MO2-klf2b
Anatomical Terms:	endocardium lateral dorsal aorta myocardium subintestinal vein
Stage Range:	Long-pec to Protruding-mouth

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	ccm2^m201/m201; s843Tg ccm2^m201/m201; s843Tg + MO1-klf2a + MO2-klf2b
Knockdown Reagents:	MO1-klf2a MO2-klf2b
Observed In:	atrium cardiac ventricle endocardial cushion endocardium heart lateral dorsal aorta endothelial cell subintestinal vein
Stage Range:	Long-pec to Protruding-mouth

Phenotype Detail

Acknowledgments

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Reprinted from Developmental Cell, 32, Renz, M., Otten, C., Faurobert, E., Rudolph, F., Zhu, Y., Boulday, G., Duchene, J., Mickoleit, M., Dietrich, A.C., Ramspacher, C., Steed, E., Manet-Dupé, S., Benz, A., Hassel, D., Vermot, J., Huisken, J., Tournier-Lasserve, E., Felbor, U., Sure, U., Albiges-Rizo, C., Abdelilah-Seyfried, S., Regulation of β1 Integrin-Klf2-Mediated Angiogenesis by CCM Proteins, 181-190, Copyright (2015) with permission from Elsevier. Full text @ Dev. Cell