Figure 2

AIP interacts with CARMA1 after T cell stimulation. (A, B) Interaction of CARMA1 and AIP in Jurkat T cells after P/I stimulation. (A) Cells were left untreated or P/I treated as indicated prior to anti-CARMA1 or IgG control IP, respectively. Co-IP was analyzed by Western Blotting. (B) Jurkat T cells were treated as in (A) and after lysis subjected to anti-AIP or IgG control IP. (C) Transient interaction of CARMA1 and AIP after CD3/CD28 co-stimulation. Jurkat T cells were stimulated as indicated prior to anti-AIP IP and analyzed by Western Blotting. (D) AIP – CBM complex association requires CARMA1. Jurkat T cells or CARMA1 deficient JPM50.6 T cells were stimulated with P/I as indicated and subjected to anti-BCL10 IP. Western Blots were stained for AIP co-precipitation. (E) AIP and BCL10 show similar binding kinetics to CARMA1 after T cell stimulation. Jurkat T cells were in parallel stimulated with P/I as indicated and after lysis subjected to anti-AIP or BCL10 IP, respectively. Western Blots were stained for CARMA1 co-precipitation. (F) Transient interaction of CARMA1 and AIP in primary mouse CD4 T cells. Cells were left untreated or stimulated with P/I as indicated prior to anti-CARMA1 IP. Co-IP was analyzed by Western Blotting.