Figure 8.

Anomanolide C inhibits TNBC cell migration in vitro by causing autophagy-dependent ferroptosis. (A-B) MDA-MB-231 cells were treated for 24 hours with AC (1 µM) alone or in conjunction with 3-MA (1 mM). 3-MA was introduced 6 hours prior to the AC treatment. The ability of the cells to migrate was subsequently evaluated utilizing the transwell assay and scratch test. Representative images and date are shown. Scale bar, 100 µm. (C-D) MDA-MB-231 cells were treated for 24 hours with AC (1 µM) alone or in conjunction with Fer-1 (1 µM). Fer-1 was added 6 hours before AC treatment. The ability of the cells to migrate was then evaluated using the transwell assay and scratch test. Representative images and date are shown. Scale bar, 100 µm. (E-F) MDA-MB-231 cells were transfected with negative-control or ATG5 siRNA. The ability of the cells to migrate was evaluated using the scratch assay and transwell assay after co-treatment with or without AC and Fer-1. Representative images and date are shown. Scale bar, 100 µm. (G-H) MDA-MB-231 cells were transfected with negative-control or overexpression of GPX4. The ability of the cells to migrate was evaluated using the scratch assay and transwell assay after co-treatment with or without AC and 3-MA. Representative images and date are shown. Scale bar, 100 µm. Data are expressed as the mean ± SEM. These results are consistent with those of at least three different experiments. ns, not significant, *, P < 0.05, **, P < 0.01, ***, P < 0.001. Statistical significance was compared between the respective AC groups.