Hesperetin (Hst) restored
palmitic acid (PA)-impaired mitophagy-mediated
degradation in HepG2 cells. Cells were preincubated with Hst for 4
h at a concentration of 40 μM and then treated with or without
PA (400 μM). (A) Representative immunoblotting and (B,C) quantification
of the indicated proteins. α-Tubulin was used as a loading control.
Cells were immunostained for LC3 (green). (D) Representative images
and (E) quantification of the numbers of punctate LC3+ structures
per cell. Representative immunostained images of (F) Tom 20 and PINK1,
(G) Tom 20 and Parkin in HepG2 cells. Nuclei were counterstained with
DAPI (blue). Data are presented as mean ± SEM of n = 3 independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 (unpaired
two-tailed student’s t test). Scale bars are
5 μm in (D), and 10 μm in (F,G).