Fig 2: Perm1 knockout downregulated fat and carbohydrate metabolism-related proteins. (a) Expression levels of fat metabolism genes in Perm1 KO mice. Expression levels of the indicated fat metabolism genes were examined by RT-PCR. Samples were normalized by 15S gene expression, including Cpt1β, Cpt2, Mcad, Lcad, and Vlcad. The mean value from WT mice was expressed as 1 (N = 6), WT (3 male + 3 female); KO (3 male + 3 female), 2 months old. (b) Expression levels of carbohydrate metabolism genes in Perm1 KO mice. Expression levels of the indicated fat metabolism genes were examined by RT-PCR. Samples were normalized by 15S gene expression, including Slc2a1, Slc2a4, Hk1, Hk2, Pdk4, and Aldoa. The mean value from WT mice was expressed as 1 (N = 6), WT (3 male + 3 female); KO (3 male + 3 female), 2 months old. (c) Western blots showing CPT1β, VLCAD, and MCAD protein levels. The intensity was normalized by GAPDH (N = 6), WT (3 male + 3 female); KO (3 male + 3 female), 2 months old. (d) The heart lysate of WT and Perm1 KO mice were analyzed with a fatty acid oxidation (FAO) assay kit. The level of FAO was normalized by that in WT mice. WT (4 male + 4 female); KO (4 male + 4 female), 2 months old. (e) Western blots showing HK2, GLUT1, and GLUT4 protein levels. The intensity was normalized by GAPDH (N = 6), WT (3 male + 3 female); KO (3 male + 3 female), 2 months old. The statistical significance was determined with Student’s t test, *p < 0.05 was considered significant.

Fig 3: SIRT3-SUMO induces Treg differentiation dependent on CPT1/VLCAD-mediated FAO. A-C qRT-PCR was employed to detect the expressions of FOXP3, CPT1, and VLCAD at various time points. D-E The expression of CPT1 was detected. G The expressions of FOXP3 and IL-10 were detected by qRT-PCR. The above data are presented as the mean ± standard deviation, *p < 0.05, n = 5

Fig 4: Monounsaturated fatty acid metabolism regulates oxidative phosphorylation levels in microglia‐like cells. BV2 cells were treated with OA. (A) The SCD2, TOM20, and VLCAD expression levels were examined by WB. (B) Real‐time OCR measured using a kit. (C) Lactate accumulation was detected using a lactate assay kit. (D, E) ECAR was evaluated using a kit. (F) ELISA was utilized to analyse FFA levels. (G) WB was conducted to assess PLIN2 expression level. (H) Flow cytometry analysis to observe lipid droplets. Data are presented as the mean ± SD; n = 3. Statistical analysis was performed by one‐way ANOVA with Tukey's test (for normally distributed data) or the Kruskal–Wallis test with Dunn's test (for non‐normally distributed data). *p < 0.05, **p < 0.01, ***p < 0.001 versus CON/T2D plasma/T2D plasma + NC.