Figure 6.

1-Deoxysphingolipids activate the NLRP3-inflammasome. Primary BMDM cells from (A,B) wild-type mice or (C,D) animals lacking expression of NLRP3 or PYCARD or pro-CASP1 were pretreated with 200 ng/mL LPS for 3 h before incubation with (A,B) 1 μM alkyne SA, (A–D) 1 μM doxSA mix or (A–D) 10 µM nigericin for 16 h or 90 min (nigericin). If applying cells were also pretreated with cytochalasin D (CyD) or bafilomycin A (Baf) or co-treated with 25 μg/mL FB1. Cytokines released to growth media were quantified by ELISA: (A,C) IL1B; (B,D) TNF. Mean values ± SEM are plotted. Statistical differences were calculated by (A,B) one-way or (C,D) two-way ANOVA followed by Tukey’s multiple comparison test. Adjusted P values: **** p < 0.0001; ns – not significant; (A,B) all vs. LPS-treated control, N = 3; (C,D) all vs. wt, N = 3