Murine Bone Marrow Derived macrophages are resistant to α hemolysin induced NLRP3-inflammasome activation.

<p>(A) Bone marrow-derived macrophages were cultured from the femurs of C57BL/6 mice, which were either wild type (WT), <i>Nlrp^−/−</i>, <i>Asc^−/−</i>, or <i>Casp1^−/−</i>. The macrophages were subsequently pretreated with HKSA or without HKSA for 3 hours then treated for 1 hour with Hla (1 µg/ml) as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007446#pone-0007446-g002" target="_blank">Fig. 2A</a>. Culture supernatant was removed and assayed for the presence of IL-1β using ELISA. Representative data from 3 independent experiments are shown. (B) Bone marrow-derived macrophages were cultured from the femurs of wild type C57BL/6 mice, as described in (A). The cells were then pretreated with nothing (N), LPS (100 ng/ml, L), or HKSA (1×10⁶ particles, H) for 3 hours. The cells were then treated for 1 hour with nothing (N.T.), 1 mM ATP, commercial α-hemolysin (1 µg/ml, Hla), or recombinant α-hemolysin (1 µg/ml, rHla). Cellular lysates and culture supernatants were prepared and subjected to immunoblot analysis with anti-IL-1β antibody.