Figure 1

GlcN inhibits activation of the NLRP3 inflammasome. (A,B) J774A.1 macrophages were incubated for 4 h with LPS (1 µg/ml) followed by incubation for 2 h with GlcN. Cells were then incubated with ATP (5 mM, 0.5 h), nigericin (10 µM, 0.5 h), MSU (100 µg/ml, 24 h) or infected with E. coli (30 MOI, 1 h). The IL-1β (A) and TNF-α (B) expression levels in the supernatants were measured by ELISA. (C–E) THP-1 macrophages (C,D) or PBMCs (E) were incubated for 4 h with LPS (1 µg/ml) followed by incubation for 2 h with GlcN, followed by incubation with ATP (5 mM, 0.5 h), MSU (100 µg/ml, 24 h) or palmitate-BSA (250 mM, 24 h). The IL-1β expression levels in the supernatants were measured by ELISA. (F) J774A.1 macrophages were incubated for 4 h with Pam3CSK4 (1 µg/ml) followed by incubation for 2 h with GlcN. Cells were then incubated with nigericin (10 µM) for 0.5 h. The IL-1β expression levels in the supernatants were measured by ELISA. The ELISA data are expressed as the means ± SD of separate experiments as indicated. *, **, *** and **** indicate a significant difference at the level of p < 0.05, p < 0.01, p < 0.001 and p < 0.0001, respectively, compared to NLRP3 activator-treated cells. (One-way ANOVA with Dunnett’s multiple comparisons test).