Fig. 6

Fusobacterium varium induces noncanonical Caspase 8 inflammasome activation in macrophages. Bone marrow-derived macrophages (BMDM) from healthy wild-type C57/Bl6 mice (age = 8–12 weeks) were cultured and treated with F. varium or F. prausnitzii (BMDM: bacteria = 1: 20) for 1 h. A Cytokine production in BMDM. ***p < 0.001; ns, not significant; by one-way ANOVA with Dunnett’s test. Experiments were repeated 3 times. B Immunostaining of IL-1β (red). Phalloidin (green) was used to outline BMDM. C Immunostaining of phospho P65 (pP65, red). Phalloidin (green) was used to outline BMDM. D–E Whole-cell lysis of BMDM treated with bacteria was subjected to Western blot analysis (40 μg per system). Experiments were repeated 3 times. D Expression of inflammasome markers in BMDM. E The level of inflammasome executors in BMDM. F Immunostaining of NLRP3 (red) and caspase-8 (green) to reveal the assembling of inflammasomes. G Immunostaining of GSDMD (red). Phalloidin (green) was used to outline BMDM. H Cell death was assessed with PI staining. **P < 0.01, ***P < 0.001; by one-way ANOVA with Dunnett’s test. Experiments were repeated 3 times