Om DNA generated in the bone marrow (BM), entire heart or semipurified cardiomyocytes afterNature. Author manuscript; obtainable in PMC 2014 November 15.van Berlo et al.Pageweeks of tamoxifen treatment in Kit/MCM RGFP mice (n=2). Bone marrow shows many of the DNA as having been recombined by Cre, whilst complete heart is just barely discernable, and purified cardiomyocytes show primarily no recombination given the sensitivity constraints of this assay. c, qPCR was also run to additional sensitively detect and quantify the extent of recombination, which was set relative for the recombination in bone marrow. Semipurified cardiomyocytes (CM) showed incredibly low rates. Averaged information are shown and error bars are s.e.m. of duplicate technical replicates from n=3 Kit/MCM RGFP mice. d, Schematic in the tamoxifen time course and timing of myocardial infarction (MI) in Kit/MCM RGFP mice. e, Echocardiography measured cardiac fractional shortening (FS ) was assessed in the mice following MI, which shows a reduction in cardiac ventricular performance at 1, two and four weeks after injury. The number of mice analyzed is shown inside the bars. Error bars represent the s.e.m. Each the handle and experimental groups showed an equivalent reduction in cardiac function postMI. f, Pictures of dissociated cardiomyocytes from hearts of Kit/MCM RGFP mice 4 weeks right after MI, which have been fixed and stained for sarcomeric actin antibody (red) and eGFP (green) at 2 various magnifications.NOTA-NHS ester Chemscene 1 eGFP cardiomyocyte is shown with sarcomeric patterning from the eGFP fluorescence.8-Hydroxyoctanoic acid In stock Author Manuscript Author Manuscript Author Manuscript Author ManuscriptNature.PMID:33544064 Author manuscript; accessible in PMC 2014 November 15.van Berlo et al.PageExtended Information Figure 7. Evaluation of eGFP myocytes inside the hearts of Kit/MCM RGFP mice just after isoproterenol infusioninduced injuryAuthor Manuscript Author Manuscript Author Manuscript Author Manuscripta, Schematic diagram displaying tamoxifen treatment of Kit/MCM RGFP mice amongst 7 and 14 weeks of age with isoproterenol (ISO) infusion occurring in between weeks 104. b, c, Quantitation and imaging of disassociated cardiomyocytes (separate images shown at two distinct magnifications) from the hearts of ISO injured Kit/MCM RGFP mice, which showed rare but definitive cardiomyocyte labeling. P0.05 vs RGFP, 31 eGFP cells of 395,302 counted from two hearts.Nature. Author manuscript; obtainable in PMC 2014 November 15.van Berlo et al.PageExtended Information Figure 8. Verifying the extent of eGFP cardiomyocytes by an independent laboratory from blinded histological heart samplesAuthor Manuscript Author Manuscript Author Manuscript Author ManuscriptUnprocessed cryosections and paraffin sections in the hearts of Kit/MCM RGFP mice after 8 weeks of tamoxifen were blinded and sent towards the Marb laboratory together with damaging manage sections from hearts that should not have staining. a, b, Two separate pictures from cryopreserved blocks are shown at 200x magnification in which the cryosection was processed for eGFP fluorescence (green) and actinin antibody (red) to show cardiomyocytes. The data show two regions exactly where a single eGFP myocyte is visible in a region with many hundred GFPnegative cardiomyocytes. The single eGFP cardiomyocyte is circled and also the inset box shows a greater magnification. Sections were also stained for nuclei (blue). Normally, about 1 definitive eGFP cardiomyocytes were identified per entire heart section inside the Marb laboratory, a result that may be constant together with the approxima.