FIGURE 4.

Parthenolide and Bay 11-7082 inhibit the NLRP3 inflammasome by targeting its ATPase activity. A, NLRP3 knock-out (NLRP3-KO) and NG5 macrophages were treated with ATP for the indicated periods of time (left and middle panels) or were treated with vehicle (DMSO), Bay 11-7082 (Bay; 12 μm), or parthenolide (Parth; 10 μm) for 15 min, followed by ATP (right panels) as indicated. The culture media were immunoblotted (IB) with anti-caspase-1 antibody (lower panels). The cells were lysed, and the resulting 6000-rpm cell pellets were cross-linked with disuccinimidyl suberate. The cross-linked pellets (upper panels) and cell lysates (middle panels) were immunoblotted with anti-ASC antibody. B, shown is the ATPase activity of immunoprecipitated recombinant NLRP3 and control immunoprecipitates in the presence of increasing concentrations of Bay 11-7082 or parthenolide as indicated. Procasp1, procaspase-1.