Adult cardiomyocyte renewal via deletion of Hippo pathway genes.

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Adult cardiomyocyte renewal via deletion of Hippo pathway genes. Adult cardiomyocyte renewal via deletion of Hippo pathway genes.Lats1/2 and Salv were conditionally deleted in cardiomyocytes using the inducible Myh6-Cre/Esr line via tamoxifen injection. All analyses were performed with 3-4 month stage hearts. (A) Diagram shows tamoxifen (Tam) and EdU injection scheme. de novo DNA synthesis was monitored by EdU incorporation (yellow) in cardiomyocytes (green) using the mTmG reporter line. (B) Percentage of EdU-positive cells for each genotype (n=3). EdU-positive cells were manually counted. (C) Percentage of cardiomyocyte nuclei in cell cycle (n=3). Isolated nuclei were stained with the cardiac marker cTnI and the cell cycle marker Ki-67. (D) Cardiomyocyte density. Numbers of cardiomyocytes were counted in each section (n=3). (E) Cardiomyocyte nucleation. Nuclei per cardiomyocyte was calculated for each genotype (n=3). (F) Cardiomyocyte cross-sectional area was measured and averaged for multiple sections per specimen (n=3). (G) Ploidy analysis (n=3). (H) Immunohistochemical analysis: Aurkb (red) and cardiomyocytes (eGFP, green). Arrowheads indicate Aurkb staining. (I) Quantification of Aurkb-positive cardiomyocytes (n=3). Error bars represent s.e.m. Todd Heallen et al. Development 2013;140:4683-4690 © 2013. Published by The Company of Biologists Ltd