Figure 3. LTA binds to the LRR domain of NLRP6.

(A, C−E) Indicated tagged NLRP6, NLRP3 or caspase-11 constructs were expressed in HEK293T cells, lysed and incubated with LTA or LPS. The lysates were immunoprecipitated (IP) and analyzed by immunoblotting (IB). (B) Full Western blots of LTA and LPS are shown as a reference. (F−H) BLI analysis of the interaction between biosensor-immobilized purified S-NLRP6 PYD and LTA, sLTA GPR or LPS. (I) Immortalized Nlrp6^−/− BMDMs reconstituted with WT NLRP6 or chimeric NLRP6 with NLRP3 PYD were infected with Listeria for 9 h. The cell lysates (Lysate) were subjected to immunoblotting. Blots of caspase-11 were cropped to reveal protein bands at different exposures. Results are representative of at least (A−D, F−I) three or (E) two independent experiments. LRR, leucine-rich repeat; PYD, pyrin domain; K_D, the equilibrium dissociation constant; sLTA, synthetic LTA; GPR, glycerophosphate repeat. See also Figure S4.