Figure 5.

CVBD induces ROS generation and NAC inhibits ROS generation, attenuating CVBD-mediated apoptosis. (A, B) T98G and U251 cells were exposed to various concentrations of CVBD for 24 h, and 0 μM CVBD was used as the control group. The ROS generation was observed by fluorescence microscopy using a CM-H₂DCFDA probe (mean ± SD of three independent experiments, **P < 0.01, ***P < 0.001 compared with the control group). (C, D) T98G and U251 cells were pre-treated with NAC (20 μM, 2 h) and post-treated with CVBD (120 μM) for 24 h, and the ROS generation was observed by fluorescence microscopy using CM-H₂DCFDA probe staining. (E) T98G and U251 cells were treated as indicated in (C), and the cell viability was examined by CCK-8 assay. (F, G) T98G and U251 cells were treated as indicated in (C), and apoptosis was detected by AnnexinV-FITC/PI staining and flow cytometry. (H, I) T98G and U251 cells were treated as indicated in (C), and the expression of PARP, C-PARP, C-Caspase3, and Cyto C (C) was determined by Western blotting analysis. GAPDH was used as a loading control (mean ± SD of three independent experiments, ***P < 0.001 compared with CVBD treatment alone).