(A) Representative immunoblots and quantitative analysis of whole-cell
heart homogenates for LC3II in WT and atg7 cKO mice.
N=5–6 in each group. Values are means ± S.E. *, p<0.05
using unpaired Student t test. (B,C) Representative fluorescent
images and quantitative analyses of mitophagy in Tg-Mito-Keima mice crossed with
WT or atg7 cKO mice and fed with either ND or HFD. Scale bar =
50μm. N=5–6 in each group. Values are means ± S.E. *,
p<0.05 using one way ANOVA followed by Bonferroni’s
post-hoc test. (D) Relative mitochondrial DNA
content/nuclear DNA content was greater in atg7 cKO mice with
HFD than in WT mice with HFD. N=5–6 in each group. Values are means
± S.E. *, p<0.05 using one way ANOVA followed by
Bonferroni’s post-hoc test. (E) Quantitative analyses of
LVW/TL. N=5–6 in each group. Values are means ± S.E. *,
p<0.05 using one way ANOVA followed by Bonferroni’s
post-hoc test. (F). Representative M mode tracing of
echocardiographs. Scale bars, horizontal 100 ms and vertical 2 mm. (G) Left
ventricular fractional shortening (LVFS) was evaluated by echocardiography.
N=5–6 in each group. Values are means ± S.E. *, p<0.05
using one way ANOVA followed by Bonferroni’s post-hoc
test. (H) ESPVR evaluated by PV-Loop analysis showed that atg7
cKO mice developed severe cardiac systolic dysfunction following 2 months of HFD
feeding. N=5–6 in each group. Values are means ± S.E. *,
p<0.05 using one way ANOVA followed by Bonferroni’s
post-hoc test. (I) EDPVR evaluated by PV-Loop analysis
showed that atg7 cKO mice developed severe cardiac diastolic
dysfunction following two months of HFD feeding. N=5–6 in each group.
Values are means ± S.E. *, p<0.05 using one way ANOVA followed by
Bonferroni’s post-hoc test. (J) Representative results
of flow cytometric analyses and quantitative analyses of TMRE intensity in adult
CMs isolated from the indicated group, showing that the MMP was decreased in
atg7 cKO mice fed with HFD. N=3 in each group. Values are
means ± S.E. *, p<0.05 using one way ANOVA followed by
Bonferroni’s post-hoc test. (K) Mitochondrial oxygen
consumption rate (OCR) in isolated CMs was evaluated with Sea Horse Analyzer.
N=3 in each group. Values are means ± S.E. *, p<0.05 using one way
ANOVA followed by Bonferroni’s post-hoc test. (L) Fatty
acid oxidation was evaluated with [3H]-oleic acid. Increases in fatty acid
oxidation in response to HFD feeding were abolished in atg7 cKO
mice. N=5 in each group. Values are means ± S.E. *, p<0.05 using
one way ANOVA followed by Bonferroni’s post-hoc
test.