Fig. 7. Characterization of 3KO embryos at E15. 5

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Fig. 7. Characterization of 3KO embryos at E15. 5 Fig. 7. Characterization of 3KO embryos at E15.5.(A–D) A subset of E15.5 3KO pups showed evidence of heart failure with peripheral edema (B, arrows) and large pericardial effusions (D, arrowheads). Characterization of 3KO embryos at E15.5.(A–D) A subset of E15.5 3KO pups showed evidence of heart failure with peripheral edema (B, arrows) and large pericardial effusions (D, arrowheads). (E–H) 3KO embryos and H/K/K littermates were subjected to immunofluorescent staining. (F) 3KO hearts at E15.5 had complete disruption of the transmural cell cycle gradient. Ventricular chambers were filled with troponin positive trabecular myocytes with an equalized distribution of pHH3 positive cells throughout the ventricular wall. (H) The overgrown trabecular region was filled with Nkx2-5 positive nuclei consistent with loss of cardiomyocyte cell cycle control. Compared to H/K/K littermate controls (G), 3KO nuclear to cytoplasmic ratios were significantly reduced (H). (I) Trabeculae of W/K/K littermates were developmentally advanced with mature sarcomeric structure (inset). (J) In contrast, 3KO trabeculae were notable for numerous mitotic figures and sarcomeres that were in various stages of disassembly/reassembly (arrowheads and inset). Scale bars: 1 mm (A–D), 100 µm (E,F), 25 µm (G,H), 2 µm (I,J). David S. Park et al. Biology Open 2013;bio.20135785 © 2013. Published by The Company of Biologists Ltd