Fig. 1: An optimized LC–MS-based cellular and mitochondrial acyl-CoA profiling workflow. | Nature Metabolism

Fig. 1: An optimized LC–MS-based cellular and mitochondrial acyl-CoA profiling workflow.

From: Cellular pan-chain acyl-CoA profiling reveals SLC25A42/SLC25A16 in mitochondrial CoA import and metabolism

Fig. 1: An optimized LC–MS-based cellular and mitochondrial acyl-CoA profiling workflow.The alt text for this image may have been generated using AI.

a, Schematic overview of the acyl-CoA profiling workflow. IP, immunoprecipitation; RP, reverse-phase. b, Extracted ion chromatograms showing the LC separation of pan-chain acyl-CoAs. c, Representative abundant acyl-CoAs from whole-cell lysates of human K562 cells. d, Validation of annotated acyl-CoAs by stable isotope labelling (acyl-CoAs, M+4) using [13C315N1]-pantothenate (1 mg l−1, 72 h) or MS/MS (Extended Data Fig. 1). e, Mass shift confirmation of representative acyl-CoAs. Black, the unlabelled acyl-CoAs and their natural isotopic peaks. Red, M+4-labelled acyl-CoAs and their natural isotopic peaks. f, Pan-chain acyl-CoA profiling of whole-cell or mitochondrial fraction from K562 cells expressing HA-MITO tag, highlighting compartment-specific enrichment of cytosolic and mitochondrial acyl-CoAs. N.D., not detected. Data in f are shown as mean ± s.d.; n = 3.

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