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. 2018 Mar 16;8:84. doi: 10.3389/fcimb.2018.00084

Figure 3.

Figure 3

Intracellular bacterial induces robust inflammasome activation and pyroptosis during CDI. (A) Peritoneal macrophages were infected with live or heat-killed C. difficile VPI 10463 for 6 h. Mature IL-1β and caspase-1 processing were analyzed by Western blotting. (B) The macrophages were infected with C. difficile in the presence of cytochalasin D (1 μM) for 6 h. Mature IL-1β and caspase-1 processing were analyzed by Western blotting. Peritoneal macrophages were infected with different strains of C. difficile (CCUG 37780, VPI 10463, and BAA 1805) for 6 h. (C) Pro-IL-1β and SLPs in cell lysate, and mature IL-1β, SLPs, and HMGB-1 in the supernatant of infected-macrophages were analyzed by Western blotting. (D) IL-1β production were assessed by ELISA. (E) Cell death induced by the different strains resulting in loss of membrane integrity was determined by the LDH release assay. (F) Mature IL-1β, SLPs, and HMGB-1 in the supernatant of infected-macrophages after caspase-1 inhibitor YVAD treatment were analyzed by Western blotting. (G) The LDH assay was also applied after the caspase-1 inhibitor Ac-YVAD-cmk (YVAD) treatment. Values represent the mean ± SEM (N = 3/group). *p < 0.05; ***p < 0.001. N.D., not detected.