Technology and genotyping of transgenic mice with cardiac restricted overexpression of human Bcl 2 have already been previously described. Bcl 2 transgenic mice were on mixed background and their non transgenic littermates were used as controls. Doxorubicin treatment was conducted with intraperitoneal injection of doxorubicin once per week for four weeks. Pitavastatin treatment was done with daily oral administration. All animal procedures were done with the approval of the Institutional Animal Care and Use Committee of Chiba University. Transthoracic echocardiography was executed with Vevo 660 equipped with (-)-MK 801 a MHz imaging transducer. All sessions were performed on conscious animals. Full intracellular oxidation in cultured cardiomyocyteswas assessed with 2?, 7? dichlorofluorescein fluorescence using CM H2DCFDA. Intracellular oxidative stress was supervised by measurement and microscopic observation of intracellular fluorescence intensity utilizing the Mithras LB940 as previously described. Measurements were performed for 5 samples in each class according to the manufacturers instruction. Histological detection of superoxide production was considered with DHE as previously described. To Organism evaluate DNA damage in cultured cardiomyocytes, CometAssay was conducted according to the manufacturers instruction. All through electrophoresis, whole DNA remains within the boundaries of the nucleus, although broken DNA migrates out of the nucleus in the form of a comet. Each comet was assigned a of 0 to 4, and 10-0 cells per slide and 3 slides per treatment were analyzed. Paraffin sections of the heart samples fixed in ten percent formalin were stained with an antibody against phosphorylated histone H2AX and dystrophin, to evaluate DNA damage in the heart in vivo. Western blot analysis was performed as previously described. Entire cell o-r tissue lysates were used for research, unless stated otherwise. For Rac1 subcellular localization assay,membrane and cytosolic proteins were prepared using proteoextract local membrane protein extraction package according to themanufacturers training. Certain signals were detected using enhanced chemiluminescence. The primary anti-bodies used for western blotting were as follows: phospho ATM, ATM, phoshop53, p53, Bax, cleaved caspase 3, Rac1, and actin. NADPH oxidase activity was measured as previously described. All measurements were done as triplicates in 96 well luminometer plates. How many viable cells purchase Dasatinib in-vitro was decided with trypan blue exclusion technique. For apoptosis investigation in vitro and in vivo, TUNEL labeling was done according to the manufacturers protocol. TUNEL positive cells were counted in 3 randomly selected low power fields from each culture plate, 3 meals for each group in-vitro.