Fig. 4.

BT-Br induces ferroptosis in DU145 cells. (a–f) Evaluation of the effects of BT-Br on intracellular ROS levels (a, b), the lipid ROS levels (c, d) and cell death (e, f) in DU145 or RWPE-1 cells. Here, the levels of ROS are evaluated through the fluorescent intensity of 2′,7′-dichlorofluorescein diacetate (DCFH-DA). And cell deaths were quantified by propidium iodide (PI) staining coupled with flow cytometry, after incubating cells with different concentrations of BT-Br for 48 h. (g–i) Evaluation of the effects of DFO on the ROS levels (g), lipid ROS levels (h) as well as cell death (i) induced by BT-Br in DU145 cells. (j) Images of DU145 cells treated with BT-Br or BT-Br/DFO respectively, quantified using PI via inverted microscope. The upper panels show the representative cell morphological changes in each group of cells (scale bar = 50 μm). The lower panels show the corresponding cell death in the upper panels. Cell death was analyzed by PI staining. Error bars represent means ± SD (n = 3); Fig. 4a–i: two-tailed unpaired t-test. *p < 0.05, **p < 0.01, ***p < 0.001, or ****p < 0.0001, ns, not significant.