Figure 3. Inflammasome formation is prevented in the presence of KCl, sucrose, quinidine and TEA.
(A) BALB/cJ BMDM were treated with LT (1 μg/ml) for the indicated times (min) in DMEM alone or DMEM containing 130 mM KCl or 300 mM sucrose. In the indicated control lanes, cells were primed with LPS (1 μg/ml; 2 h) and treated with 20 μM nigericin for 25 min. Caspase-1 activation and IL-18 maturation were monitored in cell lysates by Western blotting. (B) RAW264.7 cells were treated with 1 μg/ml LT in the presence or absence of 130 mM KCl or 300 mM sucrose for 2.5 h. Cells were then stained with MTT, and pictures of stained cells were taken using a light microscope. (C) BALB/cJ BMDM were pretreated with quinidine or TEA at the indicated concentrations for 25 min prior to LT treatment (1 μg/ml) for 85 min. Active caspase-1 and mature IL-18 were then detected in cell lysates by Western blotting. In all gels, NT refers to no-treatment control cells.
