The Protective Roles of ROS-Mediated Mitophagy on ¹²⁵I Seeds Radiation Induced Cell Death in HCT116 Cells

Figure 7

Accumulation of mitochondrial ROS is critical for ¹²⁵I seeds radiation induced mitophagy. (a–e) HCT116 cells were incubated with and without 5 mM of NAC for 4 hours and then either mock exposed or exposed to 2 Gy of ¹²⁵I seeds radiation. (a) Cells were stained with MitoSOX probe and then mitochondrial ROS were measured using flow cytometry. Typical example of flow cytometry analysis was shown. (b) Quantitative analysis of mitochondrial ROS. The values were derived from mean fluorescence intensity. The values are the means ± SD of three independent experiments. (c) Cells were stained by DCFH-DA probe and intracellular ROS were measured using flow cytometry. Typical example of flow cytometry analysis was shown. There are two peaks in the images which represented DCFH-DA negative peak and DCFH-DA positive peak, respectively. (d) Quantitative analysis of intracellular ROS. The values were derived from mean fluorescence intensity. (e) Representative Western blot analysis of LC3II/I is shown. (f–h). HCT116 cells were exposed to ¹²⁵I seeds radiation with or without Antimycin A (50 uM) or DPI (50 uM) for 18 hours. (f) Flow cytometry was performed to measure mitochondrial ROS with MitoSOX probe. Typical example of flow cytometry analysis was shown. (g) Quantitative analysis of mitochondrial ROS. The values were derived from mean fluorescence intensity. (h) Cells were immunostained with anti-LC3 antibody (green) and visualized by confocal microscopy. The typical confocal microscopy images are shown. The values are the means ± SD of three independent experiments. indicates a significant difference () as compared to the control group.

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