Skip to main content
. 2018 Dec 6;1(6):e201800237. doi: 10.26508/lsa.201800237

Figure 3. Caspase-11 dimerization is necessary and sufficient for auto-cleavage to the p32/p10 species, and protease activity.

Figure 3.

HEK293T cells were transiently transfected with pEF6-DmrB-Caspase-11 constructs depicted in (A) to mimic the p43 dimers and p32/p10 species of caspase-11 (actual predicted molecular weights of DmrB fusions are 46 kD and 35/10 kD, respectively). Transfected HEK293T cells were pre-incubated with the dimerizer drug, AP20187, for 30 min before substrate addition. (B) Cleavage of AcLEHD-afc by DmrB-Caspase-11 in digitonin-lysed HEK293T cells was monitored over 30 min. Data are mean of technical quadruplicates, and are representative of at least three biological replicate experiments. (C) HEK293T cells expressing DmrB-Caspase-11 were incubated with AP20187 for 30 min to induce dimerization. Cells were lysed with digitonin, and incubated with lysates from HEK293T expressing V5-GSDMD for 1 h at 37°C. Samples were precipitated and analysed using an immunoblot. Data are representative of three biological replicate experiments.