Figure 3.

Effect of PBME on H₂O₂-induced ROS generation and apoptosis in the C2C12 cells. The C2C12 cells were pretreated with 300 μg/mL of PBME for 1 h and then incubated with and without 1 mM H₂O₂ for 6 h. (A) To monitor the production of ROS, the cells were incubated at 37 °C in the dark for 20 min with new culture medium containing 10 μM of H2DCFDA. The generation of ROS was measured with a flow cytometer; (B) The cells were also stained with annexin V-FITC and propidium iodide (PI), and the percentages of apoptotic cells (annexin V⁺/PI⁻ cells) were then analyzed using flow cytometric analysis. The results are presented as the mean ± SD values obtained in three independent experiments (* p < 0.05 compared with the control group; ^# p < 0.05 compared with the H₂O₂-treated group).