Fig. 5

Role of inflammasomes in IL-1β release and control of N. caninum replication. Peritoneal macrophages were pre-treated with 100 μM zVAD-fmk (zVAD), 100 μM Ac-YVAD-CHO (YVAD), or 100 μM glyburide (Gly) before stimulation for 45 min, and after LPS priming for 3 h, the macrophages were infected with N. caninum (MOI = 3:1) for 12 h. a The NLRP3 expression and the secretion of IL-1β were determined by immunoblot analysis. b The production of IL-1β in the supernatants was measured by ELISA. c The number of N. caninum in infected macrophages was monitored after 12 h by qPCR. d With LPS priming wild-type (WT) and Nlrp3 ^−/− peritoneal macrophages that were infected with N. caninum (MOI = 3:1) for 12 h, and NLRP3 expression, secretion of IL-1β and cleavage of caspase-1 were determined by immunoblot analysis. e The production of IL-1β in the supernatants of WT and Nlrp3 ^−/− peritoneal macrophages stimulated with N. caninum was measured by ELISA. Abbreviations: C, control (with 0.2% DMSO in the inhibitory study); SN, supernatants; LYS, cell lysates; WT, wild-type; Nc, N. caninum; Z, zVAD-fmk; Y, Ac-YVAD-CHO; G, glyburide. The data are representative of three independent experiments and presented as the mean ± SD (*P < 0.05; **P < 0.01; ***P < 0.001 vs the Nc group or Nlrp3 ^−/− group vs WT group)