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. 2000 Jul 25;97(16):9156–9161. doi: 10.1073/pnas.160213497

Figure 6.

Figure 6

TRAF5, TRAF6, NIK, and IKKs are involved in BCMA-mediated NF-κB activation. (a) Effects of various dominant negative mutants on BCMA-mediated NF-κB activation. 293 cells were transfected with 0.5 μg of NF-κB reporter plasmid, 0.5 μg of BCMA expression plasmid, 1 μg of control plasmid, or expression plasmids for the indicated mutants. crmA plasmid (0.5 μg) also was added to the RIP () transfections for inhibiting apoptosis. Twenty hours after transfection, luciferase assays were performed and data were treated as described in Materials and Methods. Expression of the HA-tagged BCMA and the Flag- or Myc-tagged mutants was examined by Western blot analysis with antibodies against the HA, Flag, or Myc epitopes, respectively. TRADD(296S) and IκBα(SS/AA) are not tagged and could not be detected in these experiments. (b) BCMA interacts with TRAF5 and TRAF6 but not TRAF2. 293 cells were transfected with expression plasmids for HA-BCMA and the indicated plasmids. Transfected cell lysates were immunoprecipitated with anti-Flag antibody or control IgG. The immunoprecipitates were analyzed by Western blot with anti-HA antibody (Top). Expression of the transfected proteins was detected by Western blot analysis with anti-Flag (Middle) or anti-HA antibody (Bottom).