Fig. 5

ML385 cooperates with FIN56/RSL3 to promote the ferroptosis of AML cells. (a) MV4;11 and Kasumi-1 cells were treated with 5 µM ML385, 0.5 µM RSL3, or a combination of ML385 and RSL3 for 24 h (upper panel). In addition, cells were treated with 10 µM FIN56, 10 µM ML385, or a combination of FIN56 and ML385 for 48 h (lower panel), and the levels of lipid reactive oxygen species (ROS) were determined by flow cytometry using the C11-BODIPY 581/591 probe. (b) Histogram showing the levels of lipid ROS. (c) MV4;11 cells were treated with 0.5 µM RSL3, 5 µM ML385, or a combination of RSL3 and ML385. To visualize the effect of these compounds on cell death, cells were stained with propidium iodide (PI) and images were obtained using a fluorescence microscope. (d) MV4;11 cells were treated with 5 µM ML385, 0.5 µM RSL3, 10 µM ferroptosis inhibitor ferrostatin-1 (Fer-1), or the combination of ML385 and RSL3 with Fer-1 for 24 h, the percentage of Annexin V + cells was determined by flow cytometry. Data are expressed as the mean ± SD. n = 3 or more independent biological replicates, presented as individual points. P value < 0.05 was considered significant (b, d, one-way ANOVA with Bonferroni post hoc test).