Figure 5: NAC (ROS inhibitor) and Z-VAD-FMK (caspase inhibitor, Z-FMK) could reverse bigelovin-induced apoptosis. | Scientific Reports

Figure 5: NAC (ROS inhibitor) and Z-VAD-FMK (caspase inhibitor, Z-FMK) could reverse bigelovin-induced apoptosis.

From: Bigelovin triggered apoptosis in colorectal cancer in vitro and in vivo via upregulating death receptor 5 and reactive oxidative species

Figure 5

(a) HT-29 and HCT 116 cells were pretreated with indicated concentrations of NAC for 1 h, then cells were treated with bigelovin for 48 h to detect cell viability by MTT assay. Left panel in (b,c), cells were pretreated with 1 mM NAC or 20 μM Z-FMK for 1 h, then 24 h bigelovin additional treatment and whole cell proteins were analyzed by Western blot using relevant antibodies. Right panel in (b,c) are quantitative data for Western blot and data were presented in mean ± SD; *p < 0.05, **p < 0.01, ***p < 0.001 vs. control at the same time point; n = 3–4. Left panel of (d), representative histogram showing the relative intracellular ROS of cells stained with CM-H2DCFDA dye detected by flow cytometry. Cells were treated with vehicle control (Red line), 1 mM NAC (Blue line), 100 μM H2O2 (Orange line), 2.8 μM bigelovin (Green line), and 1 mM NAC plus 2.8 μM bigelovin (Purple line). Right panel of (d), quantitative effects of bigelovin on ROS production in HT-29 and HCT 116 cell lines. The values are the means ± SD of 3–4 independent experiments; **p < 0.01, ***p < 0.001 vs. vehicle control; ##p < 0.01, ###p < 0.001 vs. bigelovin treatment at indicated doses (3.6 μM for HT 29 cells and 2.8 μM for HCT 116 cells).

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