FIGURE 5.

ROS generation is enhanced by the combination of PTX and GDC. A549 cells were preincubated with GDC (20 μM) for 1 h followed by PTX (20 nM) treatment for 24 h. Intracellular ROS levels were monitored by DCFDA (A) and DHE (B,C). A549 (D) and H1299. (E) A549 cells were treated with GDC (20 μM) and PTX (20 nM) for 24 h, with or without preincubation with the mitochondrial ROS scavenger, mitoTEMPO (TEM; 10 μM). Cell viability was assessed by trypan blue exclusion assay. G + P, GDC + PTX; G + P + T, GDC + PTX + TEMPO. (F) Cells were treated as in (A); total cell lysates were immunoblotted with anti-catalase, anti-UCP2, anti-SOD1, and anti-β-actin antibodies. G + P, GDC + PTX. Protein band intensities were analyzed using ImageJ. G + P, GDC + PTX. *: p < 0.05; indicates a statistically significant difference between indicated groups. All experiments were performed at least three times. n = 3 for all groups.