Figure 7.

The SMAC mimetic BV6 enhances TNF-induced procaspase-8 processing. (A) HCT116-PI3Kmut cells were pretreated or not with 10 μM BV6 for 2 h and were then stimulated with increasing concentrations of TNF for 10 min. Phosphorylation and degradation of IκBα and cIAP1 expression were analyzed by Western blotting. (B) HCT116-PI3Kmut (–) and HCT116-PI3Kmut-TRAF2_KO (KO) cells were again pretreated with 10 μM for 2 h and then stimulated overnight with 200 ng/ml of TNF, TRAIL, or Fc-CD95L. IL8 production was determined by ELISA. Shown are results of three or four independent experiments. *p < 0.05; n.s., not significant. (C) BV6-pretreated (2 h, 10 μM) HCT116-PI3Kmut (–) and HCT116-PI3Kmut-TRAF2_KO (TRAF2-KO) cells were stimulated with the indicated concentrations of TNF for 6 h in the presence of 2.5 μg/ml CHX and subjected to Western blot analysis.