Fig. 3

MSI2 deficiency promotes the ferroptosis of CRC cells in vitro. A FACS and statistical analysis of total ROS levels (H2DCFDA) in SW620 and LOVO stable cells, gray indicates the positive control. B FACS and statistical analysis of lipid-ROS levels (C11-BODIPY) in SW620 and LOVO stable cells, gray indicates the positive control. C, FACS and statistical analysis of cell death rate in SW620 and LOVO stable cells after treatment with erastin (1–10 µg/mL) for 12 h. D, FACS and statistical analysis of cell death rate in SW620 and LOVO stable cells after treatment with erastin (5 µg/mL) and Fer-1 (10 and 20 µM) for 8 h. E The cell viability was monitored by CCK-8 and treated with different concentrations of erastin for 24 h in stable SW620, LOVO and HT29 cells. F The levels of intracellular ferrous ions were determined by treatment with different concentrations of erastin for 8 h in stable SW620, LOVO and HT29 cells. G The levels of intracellular total iron were determined by treatment with different concentrations of erastin for 8 h in stable SW620, LOVO and HT29 cells. H The levels of intracellular reduced GSH were determined by treating SW620, LOVO and HT29 stable cells with different concentrations of erastin for 8 h. I Representative images of electron microscopy revealed obvious mitochondrial abnormalities in MSI2-deficient stable cells. Scale bars, 5 μm (up) and 1 μm (bottom). J Statistical analysis of the shrunken mitochondria rate in SW620, LOVO and HT29 stable cells. These results are presented as the mean ± SD values; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; A-H, J unpaired 2-tailed Student’s t test