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. 2012 Feb 1;7(2):e31005. doi: 10.1371/journal.pone.0031005

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Chen et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A–B) Double immunofluorescence staining of the sagittal sections of E10.5 embryos with anti-NFATc1 antibody (red) and anti-alpha-SMA (green) revealed no change in the number of NFATc1⁺ endothelial cells in Ezh2-cKO hearts compared with that in littermate control hearts. (C–F) Double-immunofluorescence staining of the sagittal sections of E10.5–12.5 embryos with anti-Tbx2 antibody (red) and anti-alpha-SMA (green) revealed greatly reduced Tbx2⁺:α-SMA⁺ (arrows) primary myocardial cells of the atrioventricular canal in Ezh2^fl/fl:Nkx2.5-cre+ hearts, compared with that of the littermate control hearts. Dotted lines outlined the valve primordia. The number of Tbx2⁺:α-SMA⁺ cells was counted in three randomly selected fields of Ezh2^fl/fl:Nkx2.5-cre+ and control hearts of E10.5 or E12.5 embryos. Nuclei were counterstained with DAPI, as shown in blue. G shows the statistical analysis of Tbx2⁺:α-SMA⁺ cells between these two groups. Data are presented as mean ± SD. n = 3 for each group. *, p<0.05. Magnification, 20×. Scale bar: 100 µm.