Figure 3. The NLRP3 inflammasome is required for mCRAMP activation of caspase-1 and for release of IL-1β.
A. Murine BMDM, isolated from WT or from haplotype-matched mice deficient in NLRP3, ASC or caspase-1 were differentiated in the presence of GM-CSF for 7 days, followed by priming with 100 ng/ml LPS for 4 h. MØ were treated with 5mM ATP or 20μM mCRAMP (CMP) for 2 h and supernatants harvested for quantification by ELISA. B. Supernatants and cell lysates isolated from MØ treated as in A were analyzed for active caspase-1 (p20) content using Western analysis. Results represent mean + SEM for four separate experiments. *=p<0.05, **=p<0.01, ***=p<0.001. Rx=treatment.