Figure 3. CASPASE 8 processes CYLD at aspartate 215.
(a) Alignment of CYLD amino acid sequences from multiple species. The conserved CASPASE 8 motifs are highlighted in gray shading with the potential CASPASE 8 cleavage sites indicated by a dashed line. (b) Immunoblot of HEK 293EBNA lysates 24 hours after transfection with the indicated plasmids. (c) Immunoblot of HA-CASPASE 8 co-immunoprecipitating with FLAG-CYLD-WT or FLAG-CYLD-D215A, 24 hours post-transfection of HEK 293EBNA cells. (d) Cyld−/− MEFs reconstituted with FLAG-CYLD-WT or FLAG-CYLD-D215A were subjected to anti-FLAG immunoprecipitation after TNF stimulation for the indicated times followed by anti-FLAG blotting.