Liquid chromatography–mass spectrometry analysis of differentiating MEL cells treated with labeled itaconate. MEL cultures were induced with dimethyl sulfoxide and treated with phosphate-buffered saline (PBS) or 1 mM labeled itaconate for 72 hours. Select metabolites targeted by liquid chromatography–mass spectrometry analysis are given on the y-axes. Analyses were conducted as described in the Methods section. Bars represent means ± 1 standard deviation (n = 4 biological replicates). Multiple Student t tests combined with Bonferroni analysis produced P values <.01 for all PBS vs ¹³C₅-itaconate for the metabolites (A) ¹³C₅-itaconate, (B) ¹³C₅-itaconyl-CoA, (D) succinyl-CoA, (E) propionyl-CoA, (F) propionyl-carnitine, and (G) inositol, comparisons except (C) succinate (0.024) and (H) AMP (0.034).