Figure 4.

Effect of antioxidant treatment on mitochondrial function and morphology. A–B) Cells were treated with probucol (10 µM) for 24 h and then stained with Mitosox or TMRM to determine mitochondrial ROS levels and membrane potential. Quantification of staining intensity for Mitosox (A) and TMRM (B) in the indicated groups of cells using NIH Image J software. *p<0.05 versus all non-AD groups and probucol treated AD group. (C) Representative images with TMRM staining (Scale bar = 10 µm). Mitotracker staining was used to show mitochondria. D–E) Complex I activity (D) and ATP levels (E) were measured in the indicated groups of cells with or without probucol treatment. Data are expressed as fold increase relative to vehicle-treated non-AD cybrid cells. *p<0.05 versus all non-AD groups and probucol treated AD group. F–H) Quantitative measurement of mitochondrial density (F) and average mitochondrial length (G) in indicated cell groups using NIH Image J. *p<0.05 versus all non-AD groups and probucol treated AD group. (H) Representative images of Mitotracker Red staining. The lower panels present larger images corresponding to indicated images above (Scale bar = 5µM). I–J) Quantification of immunoreactive bands for Mfn2 (I) or DLP1 (J) relative to Hsp60 in indicated cell groups with probucol or vehicle treatment using NIH Image J software. *, p<0.05 versus non-AD groups and probucol treated AD group. Data are expressed as fold increase relative to vehicle-treated non-AD cybrid cells. Representative immunoblots are shown in the lower panel. N = 5–7 cell lines/group.