Figure 3. ITA reconstitution rescues NLRP3 inflammasome tolerance in Irg1-deficient macrophages.
(A) Design of inflammasome tolerization and ITA reconstitution (added at 4 h of LPS pre-stimulation).
(B) IL-1β secretion in BMDMs stimulated and treated as in (A). ITA or other metabolites were added as indicated, with n = 3–4 experiments. The p values were calculated using 2-way ANOVA with Sidak’s test.
(C) Pro-IL-1β detection in cell lysates of BMDMs treated and stimulated as in (A). No ATP was added.
(D) PCA of global metabolic profiles of BMDMs stimulated with LPS for 24 h and reconstituted with ITA at 4 h of stimulation, with n = 4 cultures.
(E) ITA quantification in samples from (D).
(F and G) Protein detection in BMDM lysates treated and stimulated as in (A). No ATP was added in (F). Western blots are representative of n = 3 experiments. GAPDH was used as loading control.
Data represent mean ± SEM.