The development of the doxorubicin (Dox)-induced cardiomyopathy model. (A, B) Quantification of fractional shortening (A) by echocardiography (B) in mice treated with Dox or pretreated with rosuvastatin at 10 μg/kg/d (10R) or 100 μg/kg/d (100R) at 14 days post Dox treatment. For the remainder of the study, statin mice were pretreated with 100 μg/kg/d rosuvastatin. n = 6. ∗P < 0.01 vs control (Con). (C, D) Echocardiographic assessment of cardiac function left ventricular end-diastolic dimension (LVEDD) and left ventricular end-systolic dimension (LVESD), 14 days post-Dox from Control (Con), Dox, and rosuvastatin-treated mice. Data are mean ± standard error of the mean (SEM); n = 6. (E) Heart weight corrected for body weight. Data are mean ± SEM. n = 6. (F) Heart weight corrected for tibia length. Data are mean ± SEM; n = 6. (G) Representative micrographs of Masson trichrome-stained cardiac sections in long-axis view. The white-yellow dotted line represents the border between the left ventricle (LV) and the interventricular septum (IVS), and the border between the right ventricle (RV) and the IVS. (H, I) Quantification (H) of cardiomyocyte (CM) cross-sectional area within the LV as analyzed by confocal immunofluorescence microscopy (I) at 14 d post-Dox. Green = wheat germ agglutinin for extracellular matrix (ECM); red = α-actinin; blue = 4′,6-diamidino-2-phenylindole for DNA. 100-130 cardiomyocytes quantified from 3 representative images. Data are mean ± SEM; n = 3 mice/group. (J) Quantification of mRNA levels from Dox-treated and rosuvastatin-Dox-treatedmice as analyzed by real-time reverse transcriptase quantitative polymerase chain reaction 14 d post-Dox treatment, normalized to β-actin. Data are mean ± SEM; n = 4. ∗P < 0.001 vs Con. (H, I) Representative immunoblot (K) and quantification (L) in LV protein extracts from con, Dox-treated, and statin-Dox-treated mice 14 d post-Dox, employing antibodies as indicated on the left (normalized to β-actin). Data are mean ± SEM; n = 3. #P < 0.005 Dox vs statin-Dox. ANP, atrial natriuretic peptide; Bad, Bcl-2 associated agonist of cell death; BNP, brain natriuretic peptide; Cl-Casp 3, cleaved caspase-3; CytoC, cytochrome C; Gata 4, GATA binding protein 4; Mef2, myocyte enhancer factor 2; Myh6, myosin heavy chain 6; Nkx2-5, NK2 Homeobox 5. See Supplemental Appendix S2 for original Western Blots.