Fig. 4.

The TRAF2 RING domainis required for suppression of p100 processing in resting cells, and for inhibition of TNFα-induced cell death. (a) pBa-C, pBa-T2-WT (T2-WT) and pBa-T2-ΔR (T2-ΔR) cells were left untreated, were treated with mTNFα (10ng/ml) for 15 min, or were treated with the anti-LTβR antibody (0.5μg/ml) for 4 hrs. Thereafter, p100 processing, IκBα degradation and p65 phosphorylation were monitored by Western blotting. (b) pBa-C, pBa-T2-WT and pBa-T2-ΔR cells were left untreated, were treated with mTNFα (5ng/ml) plus CHX (0.2 μg/ml), or were treated with H₂O₂ (0.075 mM) for 30 hrs, at which point cell death was assessed by microscope (x200). (c, d)The indicated cells were left untreated, were treated with mTNFα/CHX (c; 5ng/ml/0.2 μg/ml), or were treated with H₂O₂ (d; 0.075 mM), and the rate of cell death was assessed 24 and 48 hrs later via the trypan blue exclusion assay. The data shown represent the mean ± SD of three experiments performed in triplicate. (**) represents p< 0.01.