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Fig. 4. Reduced number of terminally differentiated cardiomyocytes upon loss of Dkk1. A. Sections of embryos are shown stained for cardiac troponin T (red) to label cardiomyocytes and phosho-histone H3 (PH3, green) lo mark proliferating cells and counterstained with DAPI (blue) to highlight nuclei at the stages indicated. Yellow arrowheads indicate proliferating cardiomyocytes positive for cardiac troponin and phosho-histone H3. B. Cardiac troponin T positive cardiomyocytes were quantified on continuous serial sections of stained embryos. Embryos bilaterally injected with Dkk1 MO showed reduced number of cardiomyocytes at the stages indicated. C. The mitotic index (MI) as defined by the percentage of phosho-histone H3 positive cardiomyocytes is not affected upon Dkk1 MO injection. N = number of independent embryos counted. *p < 0.05, **p < 0.01, with Mann-Whitney rank sum test. Scale bar, 100 μm. |
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Fig. 9. Cardiomyocytes detach from the myocardium upon loss of Dkk1. Sections of embryos are shown stained for cardiac troponin T (red) to label cardiomyocytes and counterstained with DAPI (blue) to highlight nuclei at the stages indicated. Upon loss of Dkk1 ectopic troponin T positive cells (white arrowheads) were found in the indicated number of cases. Embryos shown are the same as analyzed in Fig. 4. Scale bar, 100 μm. |
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Supplement Fig. 3: Loss of Dkk1 does not result in apoptosis
A,B. Apoptotic cells were stained at stage 28 by TUNEL assay. Only few apoptotic cells were detected on sections as indicated by the yellow arrowhead. No difference between Control MO and Dkk1 MO injected embryos was observed. C-F. Apoptotic cells were detected by immunofluorescence staining against cleaved caspase 3 (C-Csp3) followed by serial sections. Cardiomyocytes were detected by cardiac troponin T staining (Tnnt2). Nuclei were counterstained with DAPI. Only few apoptotic cells were detected at stage 33 (C,D) or 37 (E,F) as labelled by the yellow arrowhead. Scale bar, 100 µm.
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