Extended Data Fig. 2: Dose dependent accumulation of itaconate and derivatives in macrophages.
a, b, Intracellular levels of 4OI (blue), 4EI (yellow), and itaconate (pink) after 3h (a) or 12h (b) treatment with the indicated compound in WT primary BMDMs (n = 4 cultures). Intracellular detection of DI is not amenable to the electrospray ionization LCMS method used for these experiments. c, Intracellular levels of itaconate in unstimulated and 6h LPS-stimulated WT BMDMs that were either untreated (Control, n = 3 cultures) or pre-treated for 3h prior to stimulation with 5 mM unlabeled (12C ITA-Sigma (commercially available itaconate), 12C ITA-CTRL (itaconate synthesized as a control for the labeled itaconate), n = 2 cultures) or 13C5-itaconate (13C ITA, n = 3 cultures). Exact distribution of data can be found in Supplementary Information in Source Data files. Treatment concentrations for all experiments listed in mM. All bar plots representative of mean + s.e.m., except where n < 3 only the mean is depicted. AU = arbitrary units based on mass spectrometry peak area, and is not directly comparable between experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. P values are calculated using one-way ANOVA, and exact P values are listed in Source Data.
