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. 2016 Dec 18;11:254–262. doi: 10.1016/j.redox.2016.12.010

Fig. 2.

Fig. 2.

Artesunate induces iron-dependent, ROS-mediated ferroptosis in HNC cells. (A) Change in the number of cis-sensitive HN9 cells with treated with 50 μM Arts at different treatment days. The cells received no pretreatment or pretreatment with holo-transferrin (HTF, 20 μg/mL), the iron chelator deferoxamine (DFO, 100 mM) or the antioxidant trolox (0.5 mM). The error bars represent the standard error from three independent experiments each performed in triplicate. (B) Colony forming assay for cis-resistant HNC cells treated with 50 μM Arts for 72 h. (C) Cell death was determined with annexin V and PI staining and flow cytometry. Cell viability was measured by cellular ATP levels in HN9 cells exposed to 50 μM Arts for 72 h. The cells were pretreated with HTF, DFO, or trolox. (D) Elevation of total ROS (DCF-DA) and lipid (BODIPY C11) ROS in HN9 cells exposed to 50 μM Arts for 24 h with or without necrostatin-1 (Nec-1, 20 μM), ferrostatin-1 (Fer-1, 20 μM), and trolox (0.5 mM). The error bars represent the standard error from three replicates. *P<0.01 relative to Arts alone.