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Fig. 7

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ZDB-IMAGE-210716-100
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Figures for Zhou et al., 2015
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Fig. 7 (A–C) The endocardial-myocardial space occupied by cardiac jelly is increased in Nfatc1Cre;Krit1fl/fl;Map3k3fl/+ hearts compared with Nfatc1Cre;Krit1fl/fl;Map3k3+/+ littermates at E10.5. Higher magnification images of the boxed regions are shown on the right. (D–F) Alcian blue staining for cardiac jelly is increased in Nfatc1Cre;Krit1fl/fl;Map3k3fl/+ hearts compared with Nfatc1Cre;Krit1fl/fl;Map3k3+/+ littermates at E10.5. (G–I) Versican in cardiac jelly is increased in Nfatc1Cre;Krit1fl/fl;Map3k3fl/+ hearts compared with Nfatc1Cre;Krit1fl/fl;Map3k3+/+ littermates at E10.5. (J) The ratio of the area occupied by cardiac jelly to that occupied by myocardium in the trabeculae is increased in E10.5 Nfatc1Cre;Krit1fl/fl;Map3k3fl/+ hearts compared with Nfatc1Cre;Krit1fl/fl;Map3k3+/+ littermates. N = 3 embryos; nine sections analyzed for each group. Error bars indicate SEM. (K) Immunoblot analysis of lysate derived from whole E10.5 hearts reveals higher levels of cleaved versican (DPEAAE) in Nfatc1Cre;Krit1fl/fl;Map3k3+/+ hearts compared with Nfatc1Cre;Krit1fl/fl;Map3k3fl/+ littermates. GAPDH is shown as a loading control. (L–N) qPCR of E10.5 hearts reveals normalized expression of Klf2, Klf4, KLF2/4 target genes, Nrg1, Dll4, Tmem100, Adamts4 and Adamts5 in Nfatc1Cre;Krit1fl/fl;Map3k3fl/+ hearts compared with Nfatc1Cre;Krit1fl/fl;Map3k3+/+ littermates at E10.5. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001. Scale bars represent 100 μm. Error bars indicate SEM. (O) CCM regulation of MEKK3 activity and gene expression. The CCM complex binds MEKK3 through interaction with CCM2 and blocks MEKK3 signaling (left). Loss of the CCM complex increases MEKK3-ERK5 signaling and the expression of Klf2 and Adamts5, resulting in the breakdown of cardiac jelly and reduced myocardial proliferation (right).

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Reprinted from Developmental Cell, 32, Zhou, Z., Rawnsley, D.R., Goddard, L.M., Pan, W., Cao, X.J., Jakus, Z., Zheng, H., Yang, J., Arthur, J.S., Whitehead, K.J., Li, D., Zhou, B., Garcia, B.A., Zheng, X., Kahn, M.L., The cerebral cavernous malformation pathway controls cardiac development via regulation of endocardial MEKK3 signaling and KLF expression, 168-80, Copyright (2015) with permission from Elsevier. Full text @ Dev. Cell