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Fig. 1 AA triggered ROS-mediated cell death. (A) A549 cells were treated with AA (2 mM, 4 h), then washed with PBS, the MTT (left), or ATP (right) assay was performed after 2, 8, 20 h. (B) A549 cells were treated with AA (2 mM) for indicated times, and intracellular ROS (left) or mROS (right) generation was detected with flow cytometry. TBHP served as the positive control. (C) A549 cells were treated with AA (2 mM, 2 h), and MMP was detected with JC-1 and TMRM staining, mitochondria were tracked with MitoTracker Green. (D) A549 cells were treated with AA (2 mM) for indicated times, and lipid peroxidation was detected with flow cytometry and fluorescence microscopy. (E-G) A549 cells were treated with AA (2 mM, 2 h) with or without pretreatment of catalase (2000 U) or NAC (5 mM), and (E) intracellular ROS, (F) mROS generation, or (G) lipid peroxidation was detected with flow cytometry. (H) A549 cells were treated with AA (2 mM, 2 h) with or without pretreatment of NAC (5 mM). And MMP was detected with JC-1. (I) A549 cells were treated with AA (2 mM, 4 h) with or without pretreatment of, then washed with PBS. After 20 h, the MTT (left), or ATP (right) assay was performed. Bar = 10 μm. **p < 0.01. ***p < 0.001. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)