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Fig. 3 | Parasites & Vectors

Fig. 3

From: NLRP3 inflammasome activation in murine macrophages caused by Neospora caninum infection

Fig. 3

IL-1β production and cell death induced by N. caninum depends on the parasite amount in peritoneal macrophages. Peritoneal macrophages primed with LPS were incubated with the indicated number (MOI = parasite:cell) of N. caninum tachyzoites for 12 h (a, b, e, f) or 3 h (c, d), or incubated with ATP (5 mM) for 30 min as positive control. a, c The production of IL-1β in the supernatants was measured by ELISA. b, d The secretion of the processed p20 subunit and active IL-1β (p17) and the expression of NLRP3 were detected in cell extracts (LYS) or supernatants (SN) by immunoblot. e Supernatants were harvested at 12 h p.i., and LDH release was detected to measure cell death. f At 12 h after infection with N. caninum tachyzoites (MOI = 1:1 (4), 3:1 (5), 5:1 (6), parasite:cell), peritoneal macrophages with medium only (1), with 100 ng/ml LPS treatment (2), and 1 h after treatment with 5 mM ATP (3) were stained with a LIVE/DEAD Cell Imaging Kit for 15 min and imaged at a magnification of × 10. Live (green) and dead (red) cells in each group are shown. Abbreviations: C, control; SN, supernatants; LYS, cell lysates; MOI, multiplicity of infection. The data are representative of three independent experiments and presented as the mean ± SD (*P < 0.05; **P < 0.01; ***P < 0.001 vs negative control) Scale-bars: 50 μm

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