Figure 6

Autoradiographs showing GrB induced cleavage of murinized human BNIP-2 variants. P3′ (A) and P1′ (B) differing primed site residues following the identified P4-P1 cleavage site IEAD in hBNIP-2 were mutated to their corresponding mBNIP-2 amino acids. The in vitro transcribed and translated (mutated) BNIP-2 variants were incubated with varying concentrations (ranging from 1.95 nM to 1 μM (from left to right)) of human or mouse granzyme B. Black arrows indicate BNIP-2 precursor patterns, whereas dashed and grey arrows are indicative for BNIP-2 cleavage fragments. Percentages of cleavage are shown in the progression curves of hGrB and mGrB cleavage and indicated by full and dashed lines respectively.