Fig. 5

Acod1 regulated mitochondrial ROS levels, the activation of NF-κB, and the intermediate metabolite production in the TCA cycle. a BMDMs were transfected with Acod1 siRNA or Ctrl siRNA for 36 h, Western blot was used to detect ACOD1 protein level. b Mitochondrial ROS production was measured by FCM using Mito-SOX probes. The relative level of Mito-SOX was on the right. c The mitochondrial ROS level was determined by LCM images using the Mito-SOX probe. Red fluorescence represented mitochondrial ROS level. The relative level of Mito-SOX was on the right. d BMDMs were over-expressed Acod1 and treated with C16:0 + MSU. Mitochondrial respiration (oxygen consumption rate, OCR) was measured. e Quantification of basal respiration, maximal respiration, and OCR-coupled ATP production in mitochondrial respiration (n = 5 biological replicates). f The global ATP production was detected. g Relative levels of metabolites, assessed by LC–MS/MS analysis, in BMDMs transfected pcDNA3.1 empty plasmid (Ctrl OE) and inserted Acod1 ORF into pcDNA3.1 plasmid (ACOD1 OE) for 36 h, and then treated with C16:0 + MSU for 12 h. n = 4 independent samples. NS represents P > 0.05