Fig. 1

Representative selection of imidazole and hydantoin derivatives based on their inhibitory effect in the course of RIPK1-dependent cell death. a, c Murine L929 and NIH3T3 cells were stimulated at 37 °C for 16 h with 100 ng/ml TNFα + 25 µM zVAD (TZ) in the absence or presence of different concentrations (as indicated) of unsubstituted imidazole and its derivative hydantoin, respectively. b, d For the induction of RIPK1-dependent necroptosis, human HT-29 and U937 cells were stimulated at 37 °C for 16 h with 100 ng/ml TNFα + 1 µM SMAC mimetic SM164 + 25 µM zVAD (TSZ) in the absence or presence of the indicated amounts of imidazole and hydantoin. In each case, aromatic compounds were added 30 min before the induction of necroptosis. (E-G) L929 fibroblasts were stimulated at 37 °C for 4, 8, and 16 h with 100 ng/ml TNFα + 25 µM zVAD (TZ) in the absence or presence of Brevimytal^® (e), Luminal^® (f), or Phenhydan^® (g). Each drug was added 30 min before TZ stimulation. Necroptotic cell death was quantified by FACS analysis using 7-amino-actinomycin D and phosphatidylserine accessibility (Annexin V staining) as markers. Graphs show the mean ± SEM; n = 4 independent experiments