Supporting Figures

Files in this Data Supplement:

Fig. 6. RelB heterodimerization with p50 is not affected by RelA coexpression. The indicated NF-kB dimers were subjected to immunoprecipitation with anti-p50 antibody and analyzed for associated RelA and RelB. Note that p50 overlapped with the Ig heavy chain signal. Input controls show that protein expression levels for in vitro translated RelB and p50 were comparable in the absence or presence of RelA.

Fig. 7. RelA S276A and C216S mutants interact with p50 as efficiently as WT RelA. Whole-cell extracts from RelA-deficient MEFs infected with the indicated RelA retroviruses were subjected to immunoprecipitation (IP) with anti-p50 antibody and analyzed for associated RelA. Note that p50 overlapped with the Ig heavy chain signal.

Fig. 8. RelA serine 276 is essential for inhibition of RelB/p50 DNA binding in vitro. (A) The kB probe was incubated with the indicated in vitro translated NF-kB proteins, and DNA binding was analyzed by EMSA. (B) In vitro translated NF-kB proteins were analyzed by immunoblotting.

Fig. 9. RelB knock-down by RNAi has no effect on Bcl-xL expression. (A) WT MEFs were transfected with either a siRNA oligonucleotide targeting RelB or a scrambled control. Forty-eight hours after transfection, cells were treated with TNF-a for 8 h or left untreated, then harvested and analyzed for RelB protein expression by immunoblotting. (B) WT MEFs were transfected with either a siRNA oligonucleotide targeting RelB or a scrambled control. Forty-eight hours after transfection, cells were treated with TNF-a for 8 h or left untreated, then harvested and analyzed for Bcl-xL and b-actin expression using semiquantitative RT-PCR.