Journal: Journal of Lipid Research

Article Title: LYPLAL1 rare loss-of-function variants in humans and deletion in human hepatoma cells protect against MASLD

doi: 10.1016/j.jlr.2026.101013

Figure Lengend Snippet: LYPLAL1 knockout alters the expression of metabolic regulators and mitochondrial β-oxidation processes. A: mRNA levels of transcription factors MLXIPL and PPARα without oleic acid (OA) and with 200 μM OA. B and C: mRNA levels of (B) de novo lipogenesis genes and (C) β-oxidation genes with and without 200 μM OA treatment. D: Representative histograms and relative mean fluorescence intensity (MFI) showing protein levels of major β-oxidation enzymes ACADVL, ACADM, and HADHA without oleic acid and with 200 μM OA. E: Seahorse XF assay for mitochondrial oxygen consumption rate (OCR) in vehicle or etomoxir treated HuH-7 and LYPLAL1 KO cells without oleic acid and with 200 μM OA. Port A: Etomoxir (4 μM), port B: Oligomycin (1.5 μM), port C: FCCP (0.5 μM), and port D: Rotenone/antimycin A (0.5 μM). F: Maximal oxygen consumption rate (OCR) with vehicle or etomoxir treated HuH-7 and LYPLAL1 KO cells without oleic acid and with 200 μM OA. LYP KO = LYPLAL1 KO, Eto = Etomoxir. G: Vehicle treated (<1% DMSO) and etomoxir treated (100 μM) HuH-7 and LYPLAL1 KO cells with 200 μM OA. Cells were treated with etomoxir and oleic acid for 24 h. Data represented as mean ± S.E.M. ns: P > 0.05, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001.

Article Snippet: Probes used: GAPDH (Hs02786624_g1), ELF1 (Hs01111177_m1), MLXIPL (Hs00975714_m1), PPARα (Hs00947536_m1), ACACA (Hs01046047_m1), ACLY (Hs00982738_m1), FASN (Hs01005622_m1), SCD (Hs01682761_m1), CPT1A (Hs00912671_m1), ACADL (Hs00155630_m1), ACADM (Hs00936584_m1), ACADVL (Hs00825606_g1), EHHADH (Hs00157347_m1), HADHA (Hs00426191_m1), CD36 (Hs00354519_m1), and FABP1 (Hs00155026_m1).

Techniques: Knock-Out, Expressing, Fluorescence, XF Assay

Journal: Chinese medicine

Article Title: Xinbao Pill ameliorates heart failure via regulating the SGLT1/AMPK/PPARα axis to improve myocardial fatty acid energy metabolism.

doi: 10.1186/s13020-024-00959-1

Figure Lengend Snippet: Fig. 1 XBW effectively mitigates ISO-induced myocardial hypertrophy in NRCMs and modulates the expression of enzymes involved in myocardial fatty acid metabolism. A–B In NRCMS, Western Blot detected alterations in β-MHC and ANP protein levels, indicators of ISO-induced myocardial hypertrophy, as well as CD36, CPT-1B, and ACADM, markers of cardiac FA energy metabolism. C–D ISO-induced alterations in cell surface area of NRCMs within 48 h. The cell surface area was meticulously evaluated via Rhodamine-Phalloidin staining. Scale bar: 60 μm. E The viability of NRCMs was assessed using the MTT assay after incubation with various concentrations of XBW for 24 h. F–G After ISO stimulation and drug treatment, the WB analysis assessed the protein contents of β-MHC, ANP, ACADM, CD36, and CPT-1B. H–I The alterations in cell surface area following XBW administration were quantified. Scale bar: 60 μm. Compared with Ctrl group, ###P < 0.001, ##P < 0.01, #P < 0.05, ns: no significance; compared with ISO-induced group, ***P < 0.001, **P < 0.01, *P < 0.05, ns: no significance

Article Snippet: Antibodies against α-Tubulin (11,224-1-AP), FN (15,613-1-AP), α-SMA (14,395-1-AP), ACADM (55,210- 1-AP), CPT1-1B (22,170-1-AP), NRF1 (12,482-1-AP), PGC1α (66,369-1-IG) were purchased from Proteintech Group (Wuhan, China).

Techniques: Expressing, Western Blot, Staining, MTT Assay, Incubation

Journal: Chinese medicine

Article Title: Xinbao Pill ameliorates heart failure via regulating the SGLT1/AMPK/PPARα axis to improve myocardial fatty acid energy metabolism.

doi: 10.1186/s13020-024-00959-1

Figure Lengend Snippet: Fig. 4 XBW exhibits cardioprotective effects by mitigating the pathological alterations associated with HF and modulating fatty acid metabolism in ISO-induced HF rats. A–B Immunohistochemical staining was performed to assess the expression of ANP protein. Scale bar: 30 μm. C–D WB was employed to determine the protein levels of markers associated with hypertrophy and fibrosis in cardiac tissue. E–F Immunohistochemistry was utilized to examine the expression of CPT-1B protein. Scale bar: 30 μm. G–H The ACADM, CD36, and CPT-1B protein levels were assessed using WB analysis. Compared with Normal group, ###P < 0.001, ##P < 0.01, #P < 0.05; compared with HF group, ***P < 0.001, **P < 0.01, *P < 0.05, ns: no significance

Article Snippet: Antibodies against α-Tubulin (11,224-1-AP), FN (15,613-1-AP), α-SMA (14,395-1-AP), ACADM (55,210- 1-AP), CPT1-1B (22,170-1-AP), NRF1 (12,482-1-AP), PGC1α (66,369-1-IG) were purchased from Proteintech Group (Wuhan, China).

Techniques: Immunohistochemical staining, Staining, Expressing, Immunohistochemistry

Journal: Chinese medicine

Article Title: Xinbao Pill ameliorates heart failure via regulating the SGLT1/AMPK/PPARα axis to improve myocardial fatty acid energy metabolism.

doi: 10.1186/s13020-024-00959-1

Figure Lengend Snippet: Fig. 8 SGLT1 silencing enhances the potential of XBW in attenuating myocardial pathologic changes and enhancing myocardial fatty acid energy metabolism. A The effects of XBW on alterations in the protein levels of myocardial hypertrophy markers β-MHC and ANP after SGLT1 silence. B The effect of XBW on intracellular glucose transport after silencing SGLT1 was measured by flow cytometry. C The protein levels of FN and α-SMA, which serve as biomarkers for myocardial fibrosis, were assessed via WB analysis. D–E The effect of XBW on changes in cell surface area visualized through Rhodamine-Phalloidin staining following SGLT1 interference. Scale bar: 100 μm. F NRCMs’ ATP content level. G Immunofluorescence was employed to examine PPARα expression after silencing SGLT1. Scale bar: 10 μm. H–I WB analysis was conducted to evaluate the protein expression of SGLT1, PPARα, and myocardial fatty acid energy metabolism indicators CD36, CPT-1B, and ACADM. Independent experiments were performed at least three times with similar results. Compared with Ctrl group, ###P < 0.001, ##P < 0.01, #P < 0.05; compared with ISO-induced group, ***P < 0.001, **P < 0.01, *P < 0.05

Article Snippet: Antibodies against α-Tubulin (11,224-1-AP), FN (15,613-1-AP), α-SMA (14,395-1-AP), ACADM (55,210- 1-AP), CPT1-1B (22,170-1-AP), NRF1 (12,482-1-AP), PGC1α (66,369-1-IG) were purchased from Proteintech Group (Wuhan, China).

Techniques: Flow Cytometry, Staining, Immunofluorescence, Expressing

Journal: Chinese medicine

Article Title: Xinbao Pill ameliorates heart failure via regulating the SGLT1/AMPK/PPARα axis to improve myocardial fatty acid energy metabolism.

doi: 10.1186/s13020-024-00959-1

Figure Lengend Snippet: Fig. 9 SGLT1 overexpression blocked the effectiveness of XBW in treating myocardial pathologic changes and improving cardiac fatty acid energy metabolism A The protein levels of β-MHC and ANP under SGLT1 overexpression. B The effect of XBW on intracellular glucose transport subjected to SGLT1 overexpression. C The protein levels of FN and α-SMA were assessed via WB analysis. D–E Cell surface area was determined via Rhodamine-Phalloidin staining. Scale bar: 100 μm F Cellular ATP content levels were assessed. G IF analysis disclosed alterations in PPARα protein expression. H–I The protein contents of SGLT1, PPARα, CD36, CPT-1B, and ACADM were assessed using WB analysis to investigate the impact of SGLT1 overexpression across different experimental groups. Independent experiments were performed at least three times with similar results. Compared with Ctrl group, ###P < 0.001, ##P < 0.01, #P < 0.05; compared with ISO-induced group, ***P < 0.001, **P < 0.01, *P < 0.05, ns: no significance

Article Snippet: Antibodies against α-Tubulin (11,224-1-AP), FN (15,613-1-AP), α-SMA (14,395-1-AP), ACADM (55,210- 1-AP), CPT1-1B (22,170-1-AP), NRF1 (12,482-1-AP), PGC1α (66,369-1-IG) were purchased from Proteintech Group (Wuhan, China).

Techniques: Over Expression, Staining, Expressing

Journal: bioRxiv

Article Title: Nicotinamide riboside activates renal metabolism and protects the kidney in a model of Alport syndrome

doi: 10.1101/2024.02.26.580911

Figure Lengend Snippet: ( A,B ) NR treatment increased kidney PGC-1α in both control and Alport mice. ( C-F ) Kidney CPT1α ( C,E ) and MCAD ( D,F ), key players in mitochondrial fatty acid oxidation, were reduced in Alport mice and restored by NR treatment. Ponceau S, a nonspecific protein stain, was used as a loading control. Significance was determined by one-way ANOVA with the Holm–Šídák correction for multiple comparisons. Data are expressed as the means ± SEM. Each datum represents one mouse. * P < 0.05, ** P < 0.01, **** P < 0.0001. CPT1α, carnitine palmitoyltransferase 1-α; MCAD, medium-chain acyl-coenzyme A dehydrogenase; NR, nicotinamide riboside; PGC-1α, peroxisome proliferator-activated receptor-γ coactivator 1-α; Veh, vehicle.

Article Snippet: The primary antibodies used for immunoblotting were CPT1α (Abcam, Cambridge, United Kingdom), fibronectin (Cat. No. F3648, MilliporeSigma, Burlington, MA), MCAD (Santa Cruz Biotechnology, Dallas, TX), and PGC-1α (Cat. No. AB3242, MilliporeSigma).

Techniques: Control, Staining

Journal: Journal of animal science and biotechnology

Article Title: Dietary methionine deficiency stunts growth and increases fat deposition via suppression of fatty acids transportation and hepatic catabolism in Pekin ducks.

doi: 10.1186/s40104-022-00709-z

Figure Lengend Snippet: Fig. 4 Verification of hepatic proteomic analyses in Pekin ducks by Western blot and real-time PCR. The mRNA abundances were determined by real-time PCR analysis (a) and protein expression determined by Western Blot (b) in liver; (c, d) fold change of selected proteins between real- time PCR, Western blot validation and proteomics analysis. Results are presented as means with plus error bars (standard deviation). Differences were assessed by Student’s t test (n = 6) and denoted as follows: * P < 0.05; ** P < 0.01; *** P < 0.001. Met-D, Met-deficient; Met-A, Met-adequate. FABP1 (L-FABP), Fatty acid-binding protein, liver; ACBP, Acyl-CoA-binding protein; ACSL5, Acyl coenzyme A long-chain 5 synthetase; FATP5, Fatty acid transport protein 5; LDHA, Lactate dehydrogenase A chain; FASN, Fatty acid synthase; NDUFS1, NADH:ubiquinone oxidoreductase core subunit S1; ACADM, Acyl-CoA dehydrogenase medium chain; ALB, Albumin

Article Snippet: Primary antibodies against medium-chain specific acyl-CoA dehydrogenase (ACADM) (ab92461, Abcam, 1:5000), NADH dehydrogenase (ubiquinone) Fe-S protein 1 (NDUFS1) (ab169540, Abcam, 1:5000), albumin (ALB) (AP21444SU-N, OriGene,1:2500), lactate dehydrogenase (LDH) (WL03271, Wanleibio,1:2000), fatty acid-binding protein 1 (FABP1) (A5311, Abclonal, 1:2500), fatty acid synthase (FASN) (A0461, Abclonal, 1:2500), adipose triacylglyceride lipase (ATGL) (A6245, Abclonal, 1:2500), hormone-sensitive lipase (HSL) (A15686, Abclonal, 1: 2500), and phosphor-hormone-sensitive lipase at Ser563 (p-HSL-S563) (AP0851, Abclonal, 1:2500) were used.

Techniques: Western Blot, Real-time Polymerase Chain Reaction, Expressing, Biomarker Discovery, Standard Deviation, Binding Assay

Journal: Journal of animal science and biotechnology

Article Title: Dietary methionine deficiency stunts growth and increases fat deposition via suppression of fatty acids transportation and hepatic catabolism in Pekin ducks.

doi: 10.1186/s40104-022-00709-z

Figure Lengend Snippet: Fig. 7 Summary of the differentially expressed proteins involved in lipid metabolism induced by dietary Met deficiency. Proteins involved in lipid metabolism that were affected by dietary Met deficiency; those in red were up-expressed, whereas those in green were down-expressed. FAs, Fatty acids; TG, Triglyceride; ALB, Albumin; FA-CoA, Acyl-CoA; A-CoA, Acetyl-CoA; KB, Ketone body; PA, Pyruvic acid; LD, Lactic acid; GLU, glucose; NEFA, Non-esterified fatty acid; LDH, Lactic dehydrogenase; L-FABP (FABP1), Fatty acid-binding protein, liver; ACBP, Acyl-CoA-binding protein; ACSL5, Acyl coenzyme A long-chain 5 synthetase; FATP5, Fatty acid transport protein 5; ATGL, Adipose triacylglyceride lipase; ACADM, Acyl-CoA dehydrogenase medium chain; NDUFS1, NADH:ubiquinone oxidoreductase core subunit S1; HSL, Hormone-sensitive lipase; ATP, Adenosine Triphosphate; MDH, Malate dehydrogenase; IDH1, Isocitrate dehydrogenase; DLD, Dihydrolipoyl dehydrogenase; HADH, Hydroxyacyl-CoA dehydrogenase; GAPDH, Glyceraldehyde-3-phosphate dehydrogenase; PGK1, Phosphoglycerate kinase; HMGCS2, 3-hydroxy-3-methylglutaryl coenzyme A synthase

Article Snippet: Primary antibodies against medium-chain specific acyl-CoA dehydrogenase (ACADM) (ab92461, Abcam, 1:5000), NADH dehydrogenase (ubiquinone) Fe-S protein 1 (NDUFS1) (ab169540, Abcam, 1:5000), albumin (ALB) (AP21444SU-N, OriGene,1:2500), lactate dehydrogenase (LDH) (WL03271, Wanleibio,1:2000), fatty acid-binding protein 1 (FABP1) (A5311, Abclonal, 1:2500), fatty acid synthase (FASN) (A0461, Abclonal, 1:2500), adipose triacylglyceride lipase (ATGL) (A6245, Abclonal, 1:2500), hormone-sensitive lipase (HSL) (A15686, Abclonal, 1: 2500), and phosphor-hormone-sensitive lipase at Ser563 (p-HSL-S563) (AP0851, Abclonal, 1:2500) were used.

Techniques: Binding Assay

Journal: Nature Metabolism

Article Title: Interaction of sortilin with apolipoprotein E3 enables neurons to use long-chain fatty acids as alternative metabolic fuel

doi: 10.1038/s42255-025-01389-5

Figure Lengend Snippet: Cortical gene expression patterns were studied in male mice (12 weeks of age) of the indicated Sort1 and human APOE genotypes using quantitative RT-PCR (n = 8-18 animals per group, depending on the gene analyzed). Individual data points as well as the mean ± s.d. of the entire genotype group are given. Statistical significance of data was tested using two-way ANOVA with Tukey’s multiple comparison test. Acadl , acyl-CoA dehydrogenase long chain; Acadm , acyl-CoA dehydrogenase medium chain; Acadvl , acyl-CoA dehydrogenase very long chain; Acsl5 , acyl-CoA synthetase long chain family member 5; Cd36 , fatty acid translocase; Cpt1a , carnitine O -palmitoyltransferase 1 A; Cpt1c , carnitine O -palmitoyltransferase 1 C; Cpt2 , carnitine O -palmitoyltransferase 2; Crat , carnitine O -acyltransferase; Slc16a1 , solute carrier family 16 member 1 encoding monocarboxylate transporter (MCT) 1; Slc16a3 , solute carrier family 16 member 3 encoding MCT4; Slc16a7 , solute carrier family 16 member 7 encoding MCT2; Slc22a5 , solute carrier family 22 member 5; Slc27a1 , solute carrier family 27 member 1encoding fatty acid transport protein 1 (FATP1); Slc27a6 , solute carrier family 27 member 6, encoding FATP6.

Article Snippet: Gene expression analyses were performed using the following probes: Acadl (Mm00599660_m1), CRAT (Mm00483985_m1), Cpt1a (Mm-01231183_m1), Cpt2 (Mm00487202_m1), Acadvl (Mm00444293_m1), Acadm (Mm00431611_m1), Slc16a1 (Mm01306379_m1) and Slc22a5 (Mm00441468_m1).

Techniques: Gene Expression, Quantitative RT-PCR, Comparison

Journal: Nature Metabolism

Article Title: Interaction of sortilin with apolipoprotein E3 enables neurons to use long-chain fatty acids as alternative metabolic fuel

doi: 10.1038/s42255-025-01389-5

Figure Lengend Snippet: Gene expression patterns were compared in neurons and astrocytes isolated from the brains of male mice (12 weeks of age) of the indicated Sort1 and human APOE genotypes using fluorescence-activated cell sorting (E3WT n = 8, E3KO n = 5; E4WT and E4KO n = 7 mice). Individual data points and the mean ± s.d. of the entire genotype group are given. Statistical significance was tested using two-way ANOVA with Tukey’s multiple-comparison test. Each cell preparation was performed as an independent experiment. a , b , RT–qPCR analyses of neuronal markers NeuN and Baf53b ( a ) as well as astrocyte-specific markers Aldh1l1 and Gfap ( b ) confirm the purity of sorted neuronal and astrocyte cell preparations. c , RT–qPCR analyses of genes encoding enzymes in transport and metabolism of fatty acids document interaction of APOE and Sort1 genotypes in neurons but not astrocytes. Acadl , acyl-CoA dehydrogenase long chain; Acadm , acyl-CoA dehydrogenase medium chain; Acadvl , acyl-CoA dehydrogenase very long chain; Cpt1a , carnitine O -palmitoyltransferase 1A; Cpt2 , carnitine O -palmitoyltransferase 2; Crat , carnitine O -acyltransferase; Slc16a1 , solute carrier family 16 member 1 encoding monocarboxylate transporter (MCT) 1; Slc22a5 , solute carrier family 22 member 5.

Article Snippet: Gene expression analyses were performed using the following probes: Acadl (Mm00599660_m1), CRAT (Mm00483985_m1), Cpt1a (Mm-01231183_m1), Cpt2 (Mm00487202_m1), Acadvl (Mm00444293_m1), Acadm (Mm00431611_m1), Slc16a1 (Mm01306379_m1) and Slc22a5 (Mm00441468_m1).

Techniques: Gene Expression, Isolation, Fluorescence, FACS, Comparison, Quantitative RT-PCR