Fig 2: The expression levels of (A) p-Akt (Ser473)/t-Akt (Ser473) and p-FoxO1 (Ser256)/t-FoxO1 (Ser256), (B) p-AMPK (Thr172)/t-AMPK, PPARα, FAS, and PPARγ in the liver tissues of STZ-induced diabetic mice by oral gavage eburicoic acid (TRR). (A and B) Representative image; (C and D) quantification of the p-AMPK to t-AMPK, PPARα, FAS, and PPARγ. Protein was separated by 12% SDS-PAGE detected by western blot. All values are means ± SE (n = 8). ###p < 0.001 compared with the control (CON) group; *p < 0.05, ***p < 0.001 compared with the streptozotocin plus vehicle (distilled water) (STZ) group. TRR1, TRR2, and TRR3, eburicoic acid (TRR) (TRR1, TRR2, and TRR3, 10, 20, and 40 mg kg−1 body weight, respectively); fenofibrate (Feno, 250 mg kg−1 body weight); metformin (Metf; 300 mg kg−1 body weight).

Fig 3: circFASN overexpression blocked TAC-induced triglyceride accumulation.a Triglyceride levels in Hep-G2 cells measured by Oil red O staining in the negative control (NC), TAC only -treated, and TAC and circFASN OE plasmid-treated groups. The ratio of the area of lipid droplets to that of a single cell was used to evaluate lipid content in cells. b, c Relative expression of circFASN and miR-33a in the NC, TAC, and TAC+OE groups detected by qPCR. d Protein expression of FASN and SREBP2 in Huh7 and Hep-G2 cells analyzed by western blotting. Data represent the mean ± SD. *P < 0.05, **P < 0.01, NS, not significant, TAC groups vs. the control or OE group. The experiments were independently repeated at least three times.

Fig 4: Knockout of TLR4 in CRC reprograms cancer metabolism in vivo.A Protein expression of SREBP-1c, ACC, FASN, CPT-1, MCAD, GLUT-1, and β-catenin in the tumor tissues of mouse models inoculated with wild-type HCT116 cells or HCT116TLR4-KO cells under different dietary interventions. B–D In iTRAQ proteomics study, the gene ontology (GO) pathway enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of the biological functions of the differentially expressed proteins in the wild-type HCT116 or HCT116TLR4-KO tumor tissues dissected from HFD mice. E STRING analysis of the highlighted metabolic enzymes that show significant differences between the wild-type HCT116 or HCT116TLR4-KO tumor tissues. F ATP levels of the tumor tissues of the mouse models inoculated with wild-type HCT116 cells or HCT116TLR4-KO cells under the dietary intervention. G Quantification of the β-catenin expressions in the tumor tissues of mouse models inoculated with wild-type HCT116 cells or HCT116TLR4-KO cells under different dietary interventions. Data are shown as means ± SEM. n = 4 and 5 mice in each group. *p < 0.05, **p < 0.01 compared with CD-HCT116, a < 0.05 compared with CD-HCT116TLR4-KO, b < 0.05, bb<0.01 compared with HFD-HCT116TLR4-KO. CD-HCT116TLR4-KO mice inoculated with TLR4-KO HCT116 cells had CD diet, HFD-HCT116TLR4-KO mice inoculated with TLR4-KO HCT116 cells had HFD diet, CD-HCT116 mice inoculated with HCT116 cells had CD diet, HFD-HCT116 mice inoculated with HCT116 cells had HFD diet. ACC acetyl-CoA carboxylase, FASN fatty acid synthase, CPT1 carnitine palmitoyltransferase-1, MCAD medium-chain acyl-CoA dehydrogenase, GLUT1 glucose transporter, SREBP-1c sterol regulatory element-binding transcription factor-1.

Fig 5: The protein expressions of CD36, Fabp4, PPARγ, ACC1, and FAS in the NC, LC, NH, and LH groups determined by western blot. A The protein bands visualized by western blot. B The relative protein expression of CD36. C The relative protein expression of Fabp4. D The relative protein expression of PPARγ. E The relative protein expression of FAS. F The relative protein expression of ACC1.Data are mean ± SD(n = 3)*P < 0.05 vs NC group, #P < 0.05 vs LC group, &P < 0.05 vs NH group