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. Author manuscript; available in PMC: 2009 Feb 1.
Published in final edited form as: Cell Microbiol. 2007 Sep 10;10(2):332–343. doi: 10.1111/j.1462-5822.2007.01044.x

Figure 2. Timing of inflammasome formation in LT-sensitive BALB/cJ BMDM.

Figure 2

(A) BMDM were treated with 1 μg/ml LT for the indicated times (min). Western blot analyses were used to follow inflammasome formation, as evidenced by the appearance of the p10 fragment of active caspase-1 and mature IL-18. MEK cleavage in the same lysates was following using an N-terminal antibody whose epitope maps to the LF cleavage site of MEK1. Blots were reprobed with an antibody to EF-2 to verify equal loading. (B) BMDM were treated with LT at various concentrations for the indicated times. Culture supernatants were collected and analyzed for the presence of IL-18 by ELISA and Western blot. After supernatant removal, cells were stained with MTT to determine cell viability. (C) LPS-primed BMDM were treated with LT as in (C), and culture supernatants were analyzed for the presence of IL-1β by ELISA. Cell viability was determined by MTT staining. Filled triangles in (B) and (C) mark the LT treatments that induced the highest levels of IL-18 or IL-1β release and the cell viabilities that correspond to these treatments. In all gels, NT refers to no-treatment control cells.