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. 2020 Sep 3;9(9):820. doi: 10.3390/antiox9090820

Figure 3.

Figure 3

Mitochondrial bioenergetic disturbance and activation of mitochondrial autophagy following MG treatment in brain endothelial cells (ECs). (a,b) bEnd.3 cells were incubated with MG for 6 h and applied to Seahorse MitoStress Assay (n = 3). (a) The profile of the oxygen consumption rate was plotted. (b) The parameters for mitochondrial respiration were calculated. (c) The localization of LC3B and mitochondria was examined at indicated times after MG treatment by confocal microscopy. Mitochondria were labeled by MitoTracker. Pearson’s correlation coefficients were calculated from three independent experiments (n = 3). Representative images are shown. Scale bar: 20 μm. (d,e) The protein level of (d) Parkin-1 and (e) LC3B-II in the mitochondrial fraction was determined by western blotting. Protein levels were normalized to that of cytochrome C oxidase subunit 4 (COXIV) (n = 3). (f) Mitochondrial mass was determined by staining with NAO (n = 3). Data are presented as the mean ± SEM. * p < 0.05, ** p < 0.01 vs. control (CON).