Figure 2. Mutational analysis of Caspase-1 CARD surfaces with respect to foci formation with WT ASC CARD.
A. BiFC results of HEK293T cells transfected with either WT or mutant VN-Caspase-1 CARD together with WT VC-ASC CARD. Either lack of fluorescence or foci formation was observed for different Caspase-1 mutants. Mutants R10E, D27R, E41R, K42E, R55E, and D59R lead to loss of binding, whereas mutants R15E and R45D showed no effect on foci formation (F=Foci, N=no interaction). B. Cell lysates were immunoblotted for VN-GFP (depicting Caspase-1 CARD) and ß-actin as a loading control showing similar expression levels of mutant Caspase-1 constructs compared to WT Caspase-1.