Figure 6.

MLKL is cleaved at Asp140 upon cell death induction in MM cells. (A) Amino acid sequences alignment of the first α-helix of MLKL BR domain from multiple species. The conserved residues are highlighted with colored shading and the potential cleavage region (126-153 aa) was indicated in a rectangle with dashed border. (B) OPM2 cells were transfected with 5 µg of vectors encoding C-terminal Flag-MLKL^WT, C-terminal Flag-MLKL^D140A or C-terminal Flag-MLKL^{T357A/S358A-D140A}. After 18 h, whole cell lysates were subjected to western blotting with antibodies for C-terminus, Flag and GAPDH. (C) Cartoon diagram of MLKL structure and the cleavage by caspases. Human MLKL can be cleaved by activated caspases at Asp140 in the BR that produce a 16 kDa (N-terminus) and a 38 kDa (C-terminus) fragments.