Fig. 2

Roles of the inhibitor NAC in regulating NLRP3 inflammasome and parasite proliferation during N. caninum infection. PMs were pre-treated with or without NAC (5 mM; 1 h) and infected with N. caninum (MOI of 3:1 or 1:1, parasite: cell; 8 h). ATP (5 mM; 30 min) added to the N. caninum-infected cells was used as a positive control. At 24 h post-infection, the cell lysates and supernatants were collected. a Cell death was monitored by measuring LDH release in supernatants. b NLRP3 expression and active IL-1β (p17) was assessed by western blotting. c IL-1β production in supernatants was measured by ELISA. PMs were pre-treated with or without NAC (5 mM; 1 h) and then infected with N. caninum at a MOI of 1 for 4 h. At 24 h post-infection, the cells were collected. d Percentage of N. caninum infected cells was counted by fluorescence confocal microscopy. e Quantification of parasites in parasitophorous vacuoles were monitored by fluorescence confocal microscopy. f Number of N. caninum in infected PMs was assessed by quantitative PCR. The data are shown as the mean ± SE from three independent experiments. *P < 0.05, **P < 0.01, and ***P < 0.001 vs the NAC group