Kaplan-Meier plot of cardiac failure-free fraction and cardiac function of WT and α-MHC CaMKKβ^kd TG mice after TAC.

<p>A. α-MHC CaMKKβ^kd TG mice had a lower cardiac failure-free fraction compared with WT mice. Log-rank test indicates the significant difference between WT and α-MHC CaMKKβ^kd TG mice after TAC (p = 0.0002) (WT; n = 26, TG; n = 25). B. Representative echocardiograph pictures of WT and α-MHC CaMKKβ^kd TG hearts 3 weeks after TAC. Arrows indicate left ventricular cavities of diastolic and systolic phase. C. Serial changes in echocardiographic parameters of WT and α-MHC CaMKKβ^kd TG mice after TAC. *p<0.05 vs WT (n = 3–5 for each group).</p

Left ventricular dilatation, hypertrophy, and fibrosis is prominent in α-MHC CaMKKβ^kd TG mice compared with WT mice after TAC.

<p>A. Low-power magnifications of Masson Trichrome stained images of the left ventricle of WT and α-MHC CaMKKβ^kd TG mice with or without TAC. B. Heart weight (HW) and lung weight (LW) of WT and α-MHC CaMKKβ^kd TG mice with or without TAC. Each value was normalized to tibial length (TL). Values are the means ± S.E. of 5 mice (*p<0.05 vs sham control, †p<0.05 vs WT after TAC). C. Relative expression levels of hypertrophy-associated genes in WT and α-MHC CaMKKβ^kd TG mice with or without TAC. Values are the means ± S.E. of four mice (*p<0.05 vs sham control, †p<0.05 vs WT after TAC). D. Picrosirius staining of the left ventricle in WT and α-MHC CaMKKβ^kd TG mice with or without TAC and measurement the area of fibrosis. Values are the means ± S.E. of three to five mice (*p<0.05 vs sham control, †p<0.05 vs WT after TAC).</p

Phosphorylation of downstream targets of CaMKKβ in WT and α-MHC CaMKKβ^kd TG mice with or without TAC.

<p>Representative pictures of immunoblotting for phosphorylated form and total amount of adenosine monophosphate (AMP)-activated protein kinase (AMPK), calcium/calmodulin-dependent protein kinase (CaMK) I, and CaMKIV (A–C). Results of densitometric analysis are indicated. Values are the means ± S.E. of five mice (*p<0.05 vs sham control, †p<0.05 vs WT after TAC).</p

Generation of α-MHC CaMKKβ^kd TG mice.

<p>A. Immunoblotting and densitometry for calcium/calmodulin-dependent protein kinase kinase-β (CaMKKβ) after transverse aortic binding (TAC) in wild-type (WT) mice. Values are the means ± standard error (S.E.) of four independent experiments (*p<0.05 vs 0 week). B. Immunoblotting for CaMKKβ in WT mice and two lines of α-MHC CaMKKβ^kd TG mice. C. Immunoblotting for flag tag in two lines of α-MHC CaMKKβ^kd TG mice indicating the heart-specific overexpression of the transgene. D. Activities of CaMKKβ in the hearts of two lines of α-MHC CaMKKβ^kd TG mice by immunoprecipitate kinase assays.</p

Myocardial energy reserve measured by in situ ³¹P magnetic resonance (MR) spectroscopy.

<p>A. A ¹H MR image to define the region of interest to measure a ³¹P MR spectrum of the left ventricular anterior wall. B. In vivo cardiac ³¹P MR spectra from WT and α-MHC CaMKKβ^kd TG mice 3 weeks after TAC. C. The creatine phosphate/ATP ratio of the left ventricle in WT and α-MHC CaMKKβ^kd TG after TAC. Values are mean±standard error of the mean. *p<0.05 vs WT at same time point, n = 3–8 for each group.</p