Fig. 3.

Mitotane induces a specific form of cell death. (A) H295R cells, Jurkat T cells, HT29 cells, and HT1080 cells were treated with different doses of mitotane for 24 h. At 50 µM, the number of annexin V/7AAD double negative cells significantly decreased in the H295R cells, but all other cell lines were resistant. (B) Western blot for human phosphorylated MLKL, the downstream mediator of necroptosis. Mitotane does not induce necroptosis, as controlled by HT29 cells that were treated with TNFα, smac mimetics, and a pan-caspase inhibitor zVAD (together indicated as TSZ). GAPDH serves as a loading control. (C) Mitotane does not induce ferroptosis, as indirectly demonstrated by the absence of a protective effect of Fer-1 or Nec-1. All experiments were repeated at least 3 times; representative examples of primary data are shown.