Methods to Activate the NLRP1 Inflammasome.

In addition to being the first NLR protein proposed to form inflammasome, NLRP1s have attracted much attention in their activation mechanism by post-translational auto-proteolysis to generate C-terminal CARD containing fragment to form inflammasome. Among NLRP1, mouse NLRP1B but not human NLRP1 is well studied for its activation by lethal toxin. As dissecting the cellular components involved in NLRP1-associated diseases is highly dependent on NLRP1 inflammasome activation, experiments that can lead to NLRP1 activation is of pivotal importance to elucidate the biological role and the activation mechanism of NLRP1 especially in human. In this chapter we describe methods commonly used for mouse NLRP1B inflammasome activation as well as activation of human NLRP1 inflammasome visualized by ASC speck formation in our laboratory.