Fig. 4

ERO1α facilitates ferroptosis resistance through the upregulation of SLC7A11. A Venn diagram of the ferroptosis-related genes (FRGs) positively regulated by ERO1α and mTORC1 positively regulated genes. B sgERO1α and sgCtrl NTC/T1 null cells. C ERO1α-overexpressing Tsc2 + / + MEFs and the control cells. B and C SLC7A11 levels were examined by western blotting (left panels) and qRT–PCR (right panels). D IF analysis of the expression of SLC7A11 in the indicated cells. Scale bar, 20 μm. E–K sgERO1α NTC/T1 null cells were infected with lentiviruses carrying an empty vector (vector) or expression vectors for SLC7A11. L–R ERO1α-expressing Tsc2 + / + MEFs were infected with lentivirus harboring SLC7A11 shRNAs (shSLC7A11¹ and shSLC7A11.²) or a scrambled shRNA (shSc). E and L SLC7A11 and ERO1α protein levels were examined by western blotting. F and M Cell viability was assessed after treatment with different concentrations of erastin for 24 h in the indicated cells. G and N Representative phase-contrast images of the indicated cells treated with erastin (10 μM, 24 h) or DMSO. Scale bar, 100 μm. H–J and O–Q The indicated cells were treated with or without erastin (10 μM) for 24 h, and then L-ROS (H and O), intracellular MDA (I and P), and intracellular GSH (J and Q) were measured. K and R Representative TEM images of the indicated cells treated with 10 μM erastin for 16 h. Red arrows indicate mitochondria. Scale bar, 1 μm. Error bars indicate mean ± SD of triplicate samples. ***P < 0.001; ****P < 0.0001