CircRHOT1 reduces ferroptosis in breast cancer cells. (A) The MDA-MB-231 cells were treated with control shRNA or circRHOT1 shRNA. The expression of circRHOT1 was measured by qPCR in the cells. (B, C) The MDA-MB-231 and T47D were co-treated with 5 mmol/L erastin or ferrostatin (1 mmol/L) and circRHOT1 shRNA. The cell growth was measured by MTT assays. (D–H) The MDA-MB-231 and T47D cells were treated with control shRNA or circRHOT1 shRNA. (D) The levels of ROS were analyzed by flow cytometry analysis in the cells. (E, F) The levels of iron and Fe2+ were tested by Iron Assay Kit. (G, H) The expression of GPX4, SLC7A11, and β-actin was assessed by Western blot analysis in the cells. Data are presented as mean ± SD. Statistic significant differences were indicated: ** P < 0.01.