FIG. 11.

Effect of Bcl10 and MALT1 knockdown on the association of the ΔID with TAK1 and TRAF6 in HEK293T cells. The KD-GFP, KD-Bcl10, and KD-MALT1 cell lines were transfected with expression vectors for the wild-type CARD11 or the ΔID in the absence (−) or presence (+) of expression vectors for FLAG-tagged TAK1 (A) or TRAF6 (B). Anti-FLAG immunoprecipitations were performed as described in Materials and Methods. Where indicated, transfections included 100 ng of expression vector for wild-type CARD11 and 200 ng of expression vector for either TAK1 or TRAF6. To achieve approximately equivalent expression of the ΔID variant in each cell line, 200 ng of ΔID expression vector was used in the KD-GFP line, and 400 ng was used in the KD-Bcl10 and KD-MALT1 lines. The panels show the contents of the immunoprecipitate and lysate input developed with the indicated primary antibodies. In the control lane (C), the equivalent of 5% of the lysate input from the KD-GFP sample was resolved so as to provide a positive control for the Western blot (WB) analysis of the immunoprecipitates for the presence of Bcl10 and MALT1. WT, wild type; α, anti; IP, immunoprecipitation.