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. 2008 Jul 28;182(2):315–325. doi: 10.1083/jcb.200710067

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© 2008 Niessen et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 3. — Slug^−/− embryos display defects in AV canal EMT at E9.5. (A) Phase contrast (left) and DAPI (right) images of AV canal explants from wild-type and Slug^−/− embryos. Bars, 250 μm. (B) Quantitative analysis of EMT in AV canal explants from E9.5 wild-type (wt), Slug^+/−, and Slug^−/− embryos after 48 h in culture. Results represent the distance of a positive pixel (DAPI-stained nucleus) to the closest point of the AV canal normalized to the area of the AV canal tissue. *, P < 0.05. (C) Slug expression in the AV canal explant assay as visualized by β-galactosidase staining of wild-type and Slug-lacZ^+/− AV explants. The rounded morphology of most of the LacZ⁺ cells is shown on the right. The black square indicates the region of higher magnification shown to the right. Bars, 50 μm. (D) Representative sections of wild-type and Slug^−/− hearts counterstained with Nuclear Fast Red used for analysis in E. Dotted blue lines highlight the superior and inferior AV cushions. Bars, 50 μm. (E) Quantitation of the cellularity of the superior and inferior cushions in E9.5 wild-type (wt; n = 3) and Slug^−/− (n = 3) embryos. Error bars show SEM. (F) BrdU analysis on the percentage of proliferating cells in wild-type (n = 4) and Slug^−/− (n = 6) AV canal cardiac cushions (total), the AV canal endocardium (Endo), and AV canal mesenchymal cells (Mesen; 10–15 sections per embryo). Error bars show SEM. (G) Vector- or Slug-transduced HMEC were subjected to an endothelial wounding assay. Bars represent the distance migrated after 24 h (n = 4). *, P < 0.05. Error bars show SD. (H) Vector- or Slug-transduced HMEC were evaluated in a modified Boyden chamber assay with 20 ng/ml PDGF-BB present in the lower chamber. Bars represent the total number of cells migrated after 4 h (n = 6). *, P < 0.05. Error bars show SD.