Requirement of DAPK in functional reconstitution of NLRP3 inflammasome. Procession of mature IL-1β by reconstituted inflammasome was impaired in DAPK-knockdown HEK293T cells. The 293T cells were transduced by pLL3.7 with shDAPK or shCtrl. Control or 293T cells with DAPK knockdown were transfected with 200 ng pro-IL-1β, 25 ng NLRP3-myc, 10 ng procaspase-1, or 20 ng ASC-green fluorescent protein, as indicated. Culture supernatant and total cell lysates were prepared 48 hours after transfection. (A) The levels of IL-1β in supernatant were quantitated by ELISA. (B). The expression of DAPK, NLRP3, procaspase-1, and IL-1β in TCL and the protein levels of p17 IL-1β in the lysates were determined as described in Figure 1. **P < .01 for paired t test.