a, Chemical structures of itaconate and its derivatives.
b, Intracellular levels (n = 4 cultures) of 4OI (blue), 4EI
(yellow), and itaconate (pink) after 3h or 12h treatment with the indicated
compound in WT BMDMs. Intracellular detection of DI is not amenable to the
electrospray ionization LCMS method used for these experiments. c,
d, Intracellular levels (n = 3 cultures) of malonate (c)
and succinate (d) in WT BMDMs after 3h treatment with 5 mM malonic
acid. e, Intracellular levels of itaconate after 3h (n = 4
cultures) or 12h (n = 8 cultures) treatment with the indicated compounds in WT
BMDMs. f, Intracellular levels of itaconate after 3h or 12h
treatment (n = 3 cultures, except for WT CTRL n = 2 cultures) with the indicated
compounds followed by 24h static incubation (unstimulated) or 24h LPS
stimulation in WT or Irg1−/− BMDMs. g,
Intracellular levels of succinate after 3h (n = 4 cultures) or 12h (n = 8
cultures) treatment with the indicated compounds in WT BMDMs. h,
Intracellular levels of succinate after 3h or 12h treatment (n = 3 cultures,
except for WT CTRL n = 2 cultures) with the indicated compounds followed by 24h
static incubation (unstimulated) or 24h LPS stimulation in WT or
Irg1−/− BMDMs. Treatment
concentrations for all experiments are 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.