Figure 4.

Action of antioxidants on human gingival fibroblast-related autophagy and mitochondrial co-localization of lysosomal markers of autophagy. (A) Quantification of acidic vacuoles in control and LPS-treated fibroblasts by Lysotracker staining and flow cytometry analysis after antioxidant treatment.*P < 0.01 between control and LPS-treated fibroblasts.**P < 0.01 between LPS-treated fibroblasts and LPS + antioxidants. (B) Quantification of mitochondrial ROS in control and LPS-treated fibroblasts by MitoSOX staining and flow cytometry analysis after antioxidant treatment.*P < 0.01 between control and LPS-treated fibroblasts.**P < 0.01 between LPS-treated fibroblasts and LPS + antioxidants. (C) Protein expression of LC3 in HGF treated with LPS 10 μg/mL and antioxidants performed by western blotting as described in Methods. **P < 0.001 between LPS-treated fibroblasts and LPS + antioxidants. (D) Mitochondrial-induced ROS degraded by autophagy. Mitochondrial ROS production was localized by Mitosox Red staining. Cells were then harvested, fixed and immunostained with LC3 (autophagosome marker) and examined in a fluorescence microscope as described in Methods. Data represent the mean ± SD of three separate experiments. α-toc: α-tocopherol; BHA: butylated hydroxyanisole; CTL: control; CoQ10: coenzyme Q10; HGF: human gingival fibroblasts; LPS: lipopolysaccharide; NAC: N-acetylcysteine.

Bullon et al. BMC Medicine 2012 10:122   doi:10.1186/1741-7015-10-122
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