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. 2018 Jul 30;9:1791. doi: 10.3389/fimmu.2018.01791

Figure 2.

Figure 2

NLRP3, ASC, and caspase-1/11 control inflammasome-mediated IL-1β release in Neospora caninum infection. LPS-pre-treated bone marrow-derived macrophages (BMDMs) from wild-type (WT), Nlrp3−/−, Asc−/−, and Caspase-1/11−/− mice were infected with N. caninum at MOI = 3 for 3 or 6 h or stimulated with ATP. (A) Production of IL-1β induced by N. caninum at 3 h p.i. in supernatants was measured by ELISA. (B) Cleavage of mature IL-1β (p17) and active caspase-1 (p20) was detected by Western blotting of supernatants and cell lysates at 6 h p.i. (C) IL-1β in supernatants was measured by ELISA at 6 h p.i. BMDMs primed with LPS (100 ng/ml) or left unstimulated were infected with N. caninum at MOI = 1 for 24 h. The number of N. caninum in infected BMDMs was monitored after 24 h by quantitative PCR (D), and IL-1β in supernatants was measured by ELISA (E). Abbreviations: C, control; SN, supernatants; LYS, cell lysates; Nc, N. caninum; N.D., not detected. The data are representative of three independent experiments and are presented as the mean ± SEM (*P < 0.05, **P < 0.01, ***P < 0.001 vs. WT group).