NLRP6 and listeriosis: At the concentrations used, purified Listeriolysin O caused plasma membrane permeabilization (Fig. EV1F) but did not trigger NLRP6-dependent ASC-speck formation (Fig. 1G), indicating that Listeriolysin O promoted NLRP6 activation indirectly by allowing cytosolic entry of Listeria. Furthermore, while Listeria infection caused NLRP6 activation, it did not induce host cell permeabilization, thus excluding plasma membrane permeabilization as a potential trigger for NLRP6 activation (Fig. EV1G).