Fig. 4: IRAKM promotes IL-1β induced mitochondrial citrate transport via interaction with Slc25a1.

a Co-immunoprecipitation (IP) analysis was performed with anti-IRAKM antibody in mitochondrial lysate of IL-1β treated primary WT and IRAKM KO adipocytes and followed by western blot analysis for indicated proteins. Data are representative of three independent experiments. b Co-immunoprecipitation (IP) analysis was performed with anti-Slc25a1 antibody in mitochondrial lysate of IL-1β treated primary adipocytes and followed by western blot analysis for indicated proteins. Data are representative of three independent experiments. c Mitochondrial and cytosolic citrate levels in IL-1β stimulated WT and IRAKM KO primary adipocytes were measured by LC-MS (n = 3; mitochondria: P = 0.002; cytosol: from left to right: P = 0.023, 0.039). d Uptake rate of ¹⁴C-Citrate was measured in intact mitochondria isolated from IL-1β treated primary WT and IRAKM KO adipocytes (n = 3; from left to right: P = 0.0004, 0.0027). e Mitochondria from IL-1β treated primary adipocytes for indicated times was subjected to Lambda phosphatase (Lambda PP) treatment for 30 min, followed by western blot analysis for indicated proteins. Data are representative of three independent experiments. f Co-immunoprecipitation (IP) analysis was performed with anti-IRAKM antibody in mitochondrial lysate of IL-1β treated primary WT and IRAKM KI adipocytes and followed by western blot analysis for indicated proteins. Data are representative of three independent experiments. g Uptake rate of ¹⁴C-Citrate was measured in intact mitochondria isolated from IL-1β treated primary WT and IRAKM KI adipocytes (n = 3; from left to right: P = 0.0075, 0.03). h De novo fatty acid synthesis measured by the conversion of ¹⁴C-glucose to lipids in primary WT and IRAKM KI adipocytes treated with or without IL-1β for 24 h (n = 3; from left to right: P = 0.0006, 0.0004). i, j Flag-tagged wild-type and IRAKM Phos-mut (S167A/S170A) were restored in IRAKM KO primary adipocytes. i Co-immunoprecipitation (IP) analysis was performed with anti-Flag antibody in mitochondrial lysate of IL-1β treated primary adipocytes and followed by western blot analysis. Data are representative of three independent experiments. j De novo fatty acid synthesis measured by the conversion of ¹⁴C-glucose to lipids in primary adipocytes treated with or without IL-1β for 24 h (n = 3; from left to right: P = 0.0005, 0.001). Statistical significance was determined by two-tailed Student’s t-test (c, d, g, h, j). *P < 0.05. **P < 0.01. ***P < 0.001. All data represent mean ± s.e.m. Source data are provided as the Source data file.