Figure 5

Distribution of mitochondria generating ROS and dysfunctional mitochondria induced by GSTP silencing. (a) Observation of mitochondria membrane potential stained with JC-1 dye. Depolarized and polarized mitochondria were observed as green and red, respectively. Scale bar indicates 20 μm. Area ratios of polarized/depolarized mitochondria per cell are shown on the right. (b) Alteration of accumulated ROS in GSTP-silenced PANC-1 cells cultured with or without 50 µM NAC supplement. The amounts of ROS are represented by fluorescent intensity from the CM-H2DCFDA probe and normalized to 1 × 10⁴ cells. (c) Cell growth rates of control and GSTP-silenced PANC-1 cells cultured with or without NAC. The growth rate was normalized to 1.0 cell number on Day 0. (d) Caspase-3 and/or caspase-7 activity assay for GSTP-silenced and normal state PANC-1 cells to assess induction of apoptosis. (e) Morphological analysis in control and GSTO-silenced PANC-1 cells. Cytoskeleton protein F-actin (green) and nuclei (blue) were stained with phalloidin and DAPI, respectively. Scale bar indicates 30 μm. Measurements of mitochondria population (a), nucleus diameter and cell area (e) were performed with software ImageJ 1.51. In all figures, *and **represent P < 0.05 and P < 0.01, respectively.