Fig. 3 |. Both non-derivatized itaconate and mesaconate are immunomodulatory in macrophages.

a,b, Cytokine expression (a) and secretion (b) of RAW264.7 cells pretreated with 10 mM itaconate or mesaconate for 4 h before 3 h (a) or 21 h (b) of LPS stimulation. c,d, Cytokine expression (c) and secretion (d) of BMDMs pretreated with 10 mM itaconate or mesaconate for 4 h before 3 h (c) or 21 h (d) LPS stimulation. e, IL-1β in supernatants of BMDMs treated with indicated concentrations of mesaconate, itaconate, DMI or 4-OI, LPS stimulation as well as NLRP3 or NLRC4 inflammasome activation with nigericin (10 μM) or a mixture of BsaK (100 ng ml⁻¹) and protective antigen (1 μg ml⁻¹), respectively, following a pretreatment protocol as indicated. f, Western blots of pro-caspase-1 and cleaved caspase-1 (p20) in supernatants of BMDMs treated as in e. Data are presented as: the mean ± s.e.m. calculated from n = 12 (Ctrl, LPS), 9 (Ita1/10) or 6 (Mesa1/10) biological replicates pooled from three independent experiments (a); the mean ± s.e.m. calculated from n = 6 (IL-6) or 9 (IL-10 and TNF) biological replicates pooled from two to three independent experiments (b); the mean with overlayed data of individual mice pooled from n = 3–8 mice (each data point represent a mouse) from at least two independent experiments, and conditions from individual mice are connected with a line (c and d); the mean ± s.e.m. calculated from n = 11 mice from four independent experiments (e); representative images are from three independent experiments (f). P values were calculated by one-way ANOVA with Dunnet’s post hoc test (a, b and e) or paired t-test (paired by each mouse; c and d) and overlayed on respective comparisons.