Fig 2: Inhibiting CPT1A converts excessive FFAs into lipid drops by SOAT1.A Western blot analysis of CDK4, CDK6, CyclinD1, ACADL, and ACADM in HCC cells treated with ETO at the indicated concentration. B CCK8 assay showed that etomoxir (ETO) inhibited cell viability in the HepG2 and HUH7 (means ± SEM, n = 3); **p < 0.01, ns, no significance. C The level of total free fatty acids in HCC cells treated with CPT1A inhibition (means ± SEM, n = 3). ns: no significance. D–E Fluorescence imaging of LDs stained with BODIPY 493/503 (green) treated with pharmacological (avasimibe, 48 h) or genetic (si-RNA, 72 h) inhibition of SOAT1 in HUH7. Nuclei were stained with DAPI (blue). Scale bar, 50 µm. More than 30 cells were analyzed. Data were quantified using ImageJ software; ***p < 0.001, ****p < 0.0001, ns: no significance. F Western blot analysis of SOAT1 in HepG2 and HUH7 treated with ETO at the indicated concentration for 48 h. G–I Cellular LDs level in HCC cells treated with ETO plus AVA. More than 30 cells were analyzed.; *p < 0.05, ***p < 0.001, ****p < 0.0001.

Fig 3: AMPK is involved in the OA-mediated protein expressions of fatty acid metabolism enzymes.The gastric carcinoma cells and breast cancer cells were cultured with 0.5% BSA or 400 µM of BSA-bound oleic acid with 5 µM of Compound C or with 100 µM of AICAR for 48 h. The levels of pAMPKa (Thr172), the total AMPKa, pACC (Ser79), the total ACC, CPT1a, MCAD and ATGL were determined by western blot analysis using 50 µg of the total proteins from each sample. A representative blot is shown. (B) The cells transfected with control-siRNA or AMPKa1-siRNA (24 hours) were treated with 0.5% BSA or 400 µM of BSA-bound oleic acid for another 48 hours. The expressions of pAMPKa (Thr172), the total AMPKa, pACC (Ser79), the total ACC, CPT1a, MCAD and ATGL were determined by western blot analysis.

Fig 4: Disorder of lipid metabolism enzymes in suture-induced CNV. (A,B) Images and analysis of IHC staining of cornea paraffin sections with antibody of PPARa, CPT1A and ACADM in normal cornea and sutured cornea on day 1 to day 7 severally (brown labeling, which indicated by black arrows). Scale bar: 50 µm, n = 4 in each group. (C) Statistic analysis of qRT-PCR assay of gene expression of Ppara, Ascl1, Cpt1a and Acadm, n = 3 in each group. (D,E) Images of and statistic analysis of western blot with antibody of PPARa, ASCL1, CPT1A and ACADM in normal cornea and sutured cornea on day 1 to day 7, n = 4 in each group. ** denotes p < 0.01, one-way analysis of variance (ANOVA). Data are presented as mean ± SEM.

Fig 5: Mito-TEMPO improves the myocardial energy metabolism impaired by severe hypoglycemia in diabetic mice. (A) Representative immunoblot image of proteins related to lipid metabolism and glucose uptake. (B–G) Quantification of immunoblot image. (H) Representative immunoblot image of proteins related to oxidative phosphorylation. (I–J) Quantification of immunoblot image. (K–L) Activity of the COX I and COX IV enzymes. (M) Determination of mitochondrial potential of myocardium. (N) Determination of ATP content in cardiac tissues. Diabetic mice were treated with severe hypoglycemia for 90 min after pretreatment with MT (0.7 mg/kg/d), twice. The above evaluations were performed at the 24th hour after the termination of hypoglycemia. ##P < 0.01, ###P < 0.001 vs DM; aP<0.05, aaP<0.01, aaaP<0.001 vs DH.CD36, cluster of differentiation 36; FATP1, fatty acid transporter 1; CPT-1, carnitine palmityl transferase 1; FACS, fatty acyl coenzyme A synthetases; MCAD, medium-chain acyl-CoA dehydrogenase; GLUT4, glucose transporter 4; COX I, complex I; COX IV, complex IV; TMRE, tetramethylrhodamine ethyl ester.