Fig. 5

Functional analyses of the MALT1-BCL10 interface in Jurkat and murine CD4 T-Cells. a BCL10 KO Jurkat T-cells were lentivirally reconstituted with BCL10 wt or BM-I mutant L104R constructs. After 20 min P/I stimulation CBM complex formation was investigated by BCL10-IP. Asterisk indicates non-specific cross-reactivity of the BCL10 antibody. b, c MALT1 KO Jurkat T-cells were lentivirally reconstituted with MALT1 wt or BM-I mutant constructs V81R (b) or L82D (c). P/I stimulation and CBM complex formation was investigated as in (a). d–h NF-κB signaling (d, f, h) and MALT1 protease activation (e, g, h) in BCL10 (d, e) or MALT1 (f–h) BM-I mutant reconstituted Jurkat T-cells after P/I treatment was analyzed by Western Blot (IκBα phosphorylation and degradation) and NF-κB-DNA binding studies (EMSA). Induction of MALT1 protease activity was monitored by cleavage of MALT1 substrates in WB as indicated. i Expression of MALT1 wt and BM-I mutant (V81R) was determined by WB after enrichment of infected murine MALT1^−/− CD4 T-cells. j MALT1^−/− CD4 T-cells transduced with MALT1 wt or BM-I mutant were stimulated for 30 min with P/I. IκBα expression and degradation were measured by FACS and transduced cells were gated by co-staining of Thy1.1 (Supplementary Fig. 6a). k, l MALT1^−/− CD4 T-cells transduced as in (j) and stimulated for 5 h with P/I (k) or anti-CD3/CD28 (l). Intracellular IL-2 production was determined by FACS and transduced cells were gated by co-staining of Thy1.1 (Supplementary Fig. 6b)