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. Author manuscript; available in PMC: 2023 Jun 2.
Published in final edited form as: Nat Metab. 2022 Jun 2;4(5):524–533. doi: 10.1038/s42255-022-00565-1

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

Extended Data Fig. 3 |

a. Cytokine expression of RAW264.7 cells pre-treated with 10 mM itaconate, 0.25 mM DMI or 0.125 mM 4-OI for 4 h prior to LPS stimulation for 3 h. b. Viability of BMDMs pre-treated with indicated concentrations of mesaconate, itaconate, DMI and 4-OI for 4 h prior to LPS stimulation overnight. c. Non-targeted intracellular metabolome of RAW264.7 cells treated with 1 mM itaconate, DMI or 4-OI for 6 h. d. 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 or a mixture of BsaK and protective antigen, respectively, following a post-treatment protocol as indicated at top. Data are shown as: a,b,d. mean ± SEM calculated from (a) n = 3 biological replicates from one representative experiment, (b) n = 6 biological replicates from 3 mice, (d) n = 8 mice from 3 independent experiments; c. a representative heatmap showing z-scores of the data by row, from 2 independent experiments; P values were calculated by one-way ANOVA with Dunnet post-test (a) or paired t test (d, paired by each mouse) and overlayed on respective comparisons.