Figure 5

Inability to upregulate c-IAP2 enhances necroptotic cell death. (a and b) Indicated MEF lines were treated with mTNFα 100 ng/ml (T), BV6 2 μM (B), zVAD 20 μM (Z) and Nec-1 30 μM (N). (a) Cells were treated 20 h and viability was assessed by CellTiter-Glo. Data are mean±S.E.M. values of at least three experiments. (b and c). Total mRNA was extracted following 2 h treatment (b) or no treatment (c) and c-IAP2 mRNA levels were analyzed by quantitative RT-PCR real time. *P<0.05, **P<0.01 and ***P<0.005. (d) MEFs were transfected with the indicated siRNAs or treated with BV6 (2μM) for 48 h. Cell lysates were analyzed by western blotting with the indicated antibodies. (e) MEFs were transfected with the indicated siRNAs for 48 h. Nuclear and cytoplasmic cellular extracts were analyzed by western blotting with the indicated antibodies. The asterisk denotes a nonspecific band