Fig. 2.

Dihydrolipoamide dehydrogenase (DLD) is associated with ferroptotic cell death induced by cystine deprivation. (A–C) Silencing of DLD gene in HN3 and HN9 cell lines. PI-positive cells were stained and counted using fluorescent microscopy and flow cytometry after cystine deprivation for 48 h. Scale bar, 50 μm **P < 0.001 relative to scrambled siRNA control (siCtr). Western blotting was performed for these cells at 24 h after siDLD or siCtr transfection. (D–E) Cellular reactive oxygen species (ROS) and glutathione (GSH) contents and lipid ROS in HN3 and HN9 cells, with or without DLD gene silencing, which were subjected to CC deprivation for 8 h *P < 0.01 relative to siCtr. NS indicates statistically not significant.