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Fig. 2 AA-triggered non-apoptotic cell death. (A) A549 cells were treated with AA (2 mM, 4 h) with or without pretreatment of Z-VAD-FMK, then washed with PBS. The MTT assay was performed after 20 h. (B) A549 cells were treated with AA (2 mM, 4 h), and apoptosis was detected by Annexin V/7AAD staining. Cisplatin (CIS) (0.3 mM, 24 h) served as the positive control. (C) A549 cells were treated with AA (2 mM) for indicated times and cleaved caspase 3/7 (C-Cas 3/7) were detected. CIS (0.3 mM, 24 h) served as the positive control. (D) The cell membrane integrity of A549 cells was detected with PI staining after AA treatment. Bar = 100 μm. (E) A549 cells were treated with AA (2 mM, 4 h) with or without pretreatment of Nec-1s, GSK’872, CQ, 3-MA, DFO, Fer-1, VX765, glycine, respectively, then washed with PBS. The MTT assay was performed after 20 h. (F) A549 cells were treated with AA (2 mM) for indicated times, and P-RIP1/3, RIP1/3, p62, LC3-I/II, GPX4, CYPOR, GSDMD were detected. β-actin served as the loading control. **p < 0.01. ***p < 0.001.