(A) Evaluation of mitophagy in cardiac specific Mito-Kiema transgenic
mice fed ND or HFD for different durations. Areas with 561/457 nm ratios,
indicating mitophagy, are shown. Scale bar = 50μm. (B) Quantification of
the mitophagy area at different time points (n=8 in each group. Values are means
± S.E. *, p<0.05 using unpaired Student t test).
(C) Relative mitochondrial DNA content normalized by nuclear DNA content was
decreased after 2 months of HFD feeding (n=4 in each group. Values are means
± S.E. *, p<0.05 using unpaired Student t test).
(D,E) Representative immunoblots and quantitative analyses of whole-cell heart
homogenates and isolated mitochondria for LC3II, α-Tubulin and COXIV.
Whole-cell heart homogenates, the mitochondrial fraction, and the supernatant
fraction were prepared from mice subjected to either ND or HFD feeding for 2
months. (n=8 in each group. Values are means ± S.E. *, p<0.05
using unpaired Student t test). (F-H) Representative
fluorescent images and quantitative analyses of adult CMs isolated from cardiac
Tg-GFP-LC3 mice subjected to either ND or 2 months of HFD feeding. CMs were
incubated with Mitotracker. HFD increased either GFP-LC3 dots or GFP-LC3 rings.
Many GFP-LC3 rings were enriched with mitochondria (n=8 in each group. The
number of CMs is 50 in each group. Values are means ± S.E. *,
p<0.05 using unpaired Student t test). Scale bar = 50
μm.