Fig. 4: BioITA reports itaconate metabolism in living macrophages.

a The central nodes for cellular itaconate metabolism. SCS, succinyl-CoA ligase; AUH, methylglutaconyl-CoA hydratase; CLYBL, citramalyl-CoA lyase. b, c Representative pseudo-color images of mitochondrial (b) or cytosolic (c) BioITA in RAW264.7 cells with or without Irg1 KO were captured at different time points after LPS (10 ng ml–1) stimulation (b, c, upper panels). Time-dependent fluorescence intensity of BioITAs or dBioITA in individual cell after LPS stimulation was quantitated (b, c, lower panels). Mitochondrial or cytosolic dBioITA served as a control. Scale bar, 20 μm (b, c, upper panels). Data are shown as mean ± SD (n = 8) (b, c, lower panels). d, e Representative pseudo-color images of mitochondrial (d) or cytosolic (e) BioITA in RAW264.7 cells with or without Suclg1 KO were captured at different time points after LPS (10 ng ml–1) stimulation (d, e, upper panels). Time-dependent fluorescence intensity of BioITAs or dBioITA in individual cell after LPS stimulation was quantitated (d, e, lower panels). Mitochondrial or cytosolic dBioITA served as a control. Scale bar, 20 μm (d, e, upper panels). Data are shown as mean ± SD (n = 8) (d, e, lower panels). Dotted lines indicated the mean ± SD (b–e, lower panels). Source data are provided as a Source Data file.