Figure 8.

Bid and JNK are involved in receptor-interacting protein kinase 1/receptor-interacting protein kinase 3 (RIPK3)/mixed lineage kinase domain-like protein (MLKL) signaling, leading to complement-induced necrosis. General protocol: cells were pretreated with the indicated inhibitor for 1 h at 37°C and then treated with NHS for 1 h, at 37°C. The percentage of necrotic cell death (CD) and the percentage of CD inhibition by the inhibitor, relative to DMSO, were calculated (after data conversion to y/1 − y) as explained in Section “Materials and Methods.” *P < 0.01 relative to DMSO (A,C,D,F) or between WT and KO cells (B,D) (t-test). (A) WT, JNK1, or JNK2 mouse embryonic fibroblasts (MEFs) were pretreated with necrostatin-1 (Nec-1) (50 µM) or DMSO as control. (B) WT and RIPK3 KO MEFs were pretreated with SP600125 or DMSO as control. (C) WT and MLKL KO MEFs were treated with 10 µM SP600125 or DMSO. (D) WT or Bid KO MEFs were treated with Nec-1 or DMSO. (E) WT or Bid KO MEFs were pretreated with GSK’872 at 2 or 5 µM or with DMSO as control. (F) WT or Bid KO MEFs were treated with 1 µM GW806742X or DMSO. All results represent at least three independent experiments.