rabbit anti cept1  (Bioss)

Journal: bioRxiv

Article Title: Endothelial CEPT1 Regulates Hepatic MTTP-Mediated Lipid Metabolism and Impacts Aortic Atherosclerosis

doi: 10.1101/2025.09.21.677657

Figure Lengend Snippet: (A) Quantification of human MTTP activity at the plateau phase (170-210 min) non-steatosis (n=11) and steatosis samples (n=12). Each patient is represented by a group of approximately 5 data points (dots). (B-C) Representative western blot showing reduced CEPT1 and MTTP protein levels in human liver tissue from non-steatosis (n=11) and steatosis samples (n=12). GAPDH serves as a loading control. (D) Quantification of hepatic CEPT1 (C) and MTTP (D) protein levels. (E-F) Representative immunofluorescence images of human liver sections, comparing non-steatosis and steatosis conditions: (E) H&E staining with DAPI (blue) and MTTP (green); white arrows highlight regions of positive MTTP staining; (F) H&E staining alongside DAPI (blue), CD31 (red), and CEPT1 (green), white arrows indicate areas of CD31–CEPT1 colocalization, shown in yellow; (G) Quantification of MTTP integrated density, showing a significant decrease in steatosis (*p<0.05). (H) Quantification of CEPT1/CD31 co-localization, which is significantly reduced in steatotic samples (**p<0.01). Data represent mean ± SEM. Statistical significance was determined by Unpaired t-test.

Article Snippet: Sections were incubated overnight at 4°C with the following primary antibody anti-CD68 (1:200, Bio-Rad, MCA17576A), CD31 (1:250, Santa Cruz, sc-376764), MTTP (1:150, Thermo Fischer, PA5-76049) and CEPT1 (1:250, Bioss, bs-12284R-Cy3).

Techniques: Activity Assay, Western Blot, Control, Immunofluorescence, Staining

Journal: bioRxiv

Article Title: Endothelial CEPT1 Regulates Hepatic MTTP-Mediated Lipid Metabolism and Impacts Aortic Atherosclerosis

doi: 10.1101/2025.09.21.677657

Figure Lengend Snippet: (A) Schematic illustrating the proposed impact of endothelial CEPT1 (EC CEPT1) on hepatic MTTP and PPARα regulation. EC CEPT1 influences hepatocyte function, potentially impacting VLDL assembly and fatty acid metabolism. Fenofibrate (FEN), a PPARα activator, is included to indicate pharmacological activation of PPARα in this pathway (B) Experimental setup illustrating co-culture of HUVECs with HepG2 cells ± FEN treatment (50 µM) for 48h. HUVECs were transfected with si CEPT1 +. (C-E) Relative mRNA expression of CEPT1 (C), PPARα (D), and MTTP (E) in HUVECs transfected with siRNA targeting Cept1 compared to untransfected HUVECs (control), normalized to controls (n=3). Relative mRNA expression levels of MTTP (F) and PPARα (G) in HepG2 cells co-cultured with HUVECs ± siRNA targeting CEPT1 and ±FEN treatment (50µM) for 48h, normalized to controls (n=3). (H) MTTP activity curve (pmol transferred/µg protein) over time for HepG2 cells from control and co-culture with HUVEC si CEPT1 + cells ±FEN treatment (50µM for 48h). Plateau indicates maximal MTTP transfer capacity, as determined by endpoint assay guidelines. (I) Quantification of MTTP activity at the plateau phase (195-240 min) for HepG2 cells from control and co-culture with HUVEC si CEPT1 + cells ±FEN treatment. Data were analyzed by Mann-Whitney U test; n=4. (J) Experimental setup illustrating co-culture of HepG2 with si PPARα + transfected HUVECs vs control (HUVECs si PPARα -) for 48h. (K) Relative mRNA expression of MTTP in HepG2 co-culture with HUVECs si PPARα + vs positive control (HUVECs si GFP + ) and negative control (un-transfected HUVECs) (n=9). Data were analyzed by t-test or one-way ANOVA followed by Tukey’s multiple comparisons test. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: Sections were incubated overnight at 4°C with the following primary antibody anti-CD68 (1:200, Bio-Rad, MCA17576A), CD31 (1:250, Santa Cruz, sc-376764), MTTP (1:150, Thermo Fischer, PA5-76049) and CEPT1 (1:250, Bioss, bs-12284R-Cy3).

Techniques: Activation Assay, Co-Culture Assay, Transfection, Expressing, Control, Cell Culture, Activity Assay, End Point Assay, MANN-WHITNEY, Positive Control, Negative Control

Journal: bioRxiv

Article Title: Endothelial CEPT1 Regulates Hepatic MTTP-Mediated Lipid Metabolism and Impacts Aortic Atherosclerosis

doi: 10.1101/2025.09.21.677657

Figure Lengend Snippet: (A) Schematic representation of experimental design. Tamoxifen was administered intraperitoneally to induce Cre recombinase activity in Cept1 fl/fl Apoe +/+ and Cep1 fl/fl Apoe -/- mice. After tamoxifen treatment, mice were fed a 42% high-fat diet for 12 weeks. Serum samples were collected weekly for further analysis. (B) Body weight measurements (in grams) of mice over 12-week period. (C-E) Serum lipid profile after 12 weeks on a high-fat diet. Mean serum levels of total cholesterol (mg/dL) (C), triglycerides (mg/dL) (D), and free fatty acids (ng/dL) (E) in Cept1 fl/fl Apoe +/+ and Cep1 fl/fl Apoe -/- mice compared to controls. All data are presented as mean ± SEM. Statistical significance was determined by Unpaired t-test. *p<0.05, **p<0.01, ****p<0.0001, and n ≥ 6 for all groups.

Article Snippet: Sections were incubated overnight at 4°C with the following primary antibody anti-CD68 (1:200, Bio-Rad, MCA17576A), CD31 (1:250, Santa Cruz, sc-376764), MTTP (1:150, Thermo Fischer, PA5-76049) and CEPT1 (1:250, Bioss, bs-12284R-Cy3).

Techniques: Activity Assay

Journal: bioRxiv

Article Title: Endothelial CEPT1 Regulates Hepatic MTTP-Mediated Lipid Metabolism and Impacts Aortic Atherosclerosis

doi: 10.1101/2025.09.21.677657

Figure Lengend Snippet: (A) Representative H&E-stained liver sections from Cept1 fl/fl Cre + Apoe -/- , Cept1 fl/fl Cre - Apoe -/- , Cept1 fl/fl Cre + Apoe +/+ and Cept1 fl/fl Cre - Apoe +/+ mice. Scale bar = 100 μm. (B) Quantification of hepatic lipid accumulation as measured by ORO intensity in liver sections across all experimental groups. Data analyzed by one-way ANOVA with Tukey’s post-hoc test; n=4-6 per group; **p<0.01. (C) Representative Western blots of MTTP, PPARα, CEPT1, and GAPDH in liver lysates of Cept1 fl/fl Cre - Apoe +/+ and Cept1 fl/fl Cre + Apoe +/+ . Molecular weights (kDa) are indicated. Quantification of hepatic Cept1 fl/fl Cre - Apoe +/+ and Cept1 fl/fl Cre + Apoe +/+ MTTP (D), PPARα (E), CEPT1 (F) protein levels normalized to GAPDH in all mice groups; n=3. Hepatic Mttp (G) and Cept1 (H) gene expression in Cept1 fl/fl Cre - Apoe +/+ and Cept1 fl/fl Cre + Apoe +/+ . (I) Representative Western blots of MTTP, PPARα, CEPT1, and GAPDH in liver lysates of Cept1 fl/fl Cre - Apoe -/- and Cept1 fl/fl Cre + Apoe -/- . Molecular weights (kDa) are indicated. Quantification of hepatic Cept1 fl/fl Cre - Apoe +/+ and Cept1 fl/fl Cre + Apoe -+/+ MTTP (J), PPARα (K), CEPT1 (L) protein levels normalized to GAPDH in all mice groups; n=3. Hepatic Mttp (M) and Cept1 (N) gene expression in Cept1 fl/fl Cre - Apoe -/- and Cept1 fl/fl Cre + Apoe -/- . Data were analyzed by t-test. (O) Liver MTTP activity curve (pmol transfered/μg protein) over time in Cept1 fl/fl Cre + Apoe -/- , and Cept1 fl/fl Cre + Apoe +/+ mice compared to controls. Plateau represents the maximal transfer capacity achieved under these experimental conditions, as per manufacturer’s guidelines for endpoint assays. (P-Q) MTTP activity at the plateau phase (180-240 min) in Cept1 fl/fl Cre + Apoe +/+ (P), and Cept1 fl/fl Cre + Apoe -/- (Q) mice compared to controls. Data were analyzed by Mann-Whitney U test; n=4. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: Sections were incubated overnight at 4°C with the following primary antibody anti-CD68 (1:200, Bio-Rad, MCA17576A), CD31 (1:250, Santa Cruz, sc-376764), MTTP (1:150, Thermo Fischer, PA5-76049) and CEPT1 (1:250, Bioss, bs-12284R-Cy3).

Techniques: Staining, Western Blot, Gene Expression, Activity Assay, MANN-WHITNEY

Journal: bioRxiv

Article Title: Endothelial CEPT1 Regulates Hepatic MTTP-Mediated Lipid Metabolism and Impacts Aortic Atherosclerosis

doi: 10.1101/2025.09.21.677657

Figure Lengend Snippet: Representative images of ORO stained en face preparations of the aortas in Cept1 fl/fl Cre - Apoe +/+ and Cept1 fl/fl Cre + Apoe +/+ (A). Scale bar=5mm. Quantification of plaque area in the total aorta (B; n ≥ 5), aortic arch (C; n ≥ 4), thoracic aorta (D; n ≥ 5), and aortoiliac segment (E; n ≥ 4) of Cept1 fl/fl Cre - Apoe +/+ , and Cept1 fl/fl Cre + Apoe +/+ mice. Representative images of ORO stained en face preparations of the aortas in Cept1 fl/fl Cre - Apoe -/- and Cept1 fl/fl Cre + Apoe -/- (F). Scale bar=5mm. Quantification of plaque area in the total aorta (G; n ≥ 5), aortic arch (H; n ≥ 4), thoracic aorta (I; n ≥ 5), and aortoiliac segment (J; n ≥ 4) of Cept1 fl/fl Cre - Apoe -/- , and Cept1 fl/fl Cre + Apoe -/- mice. (K) Representative images of ORO-stained aortic root sections from Cept1 fl/fl Cre + Apoe +/+ and Cept1 fl/fl Cre + Apoe -/- mice compared to controls. Scale bar=100µm. (L-M) Quantification of lipid deposition in the aortic root sections; n ≥5 Cept1 fl/fl Cre + Apoe +/+ (L) and Cept1 fl/fl Cre + Apoe -/- (M) compared to their corresponding controls. (N) Representative immunofluorescence staining for CD68 (green) and DAPI (blue) in the aortic root of Cept1 fl/fl Cre - Apoe -/- and Cept1 fl/fl Cre + Apoe -/- mice. Scale bar=100μm. (O) Quantifying means CD68 intensity (arbitrary units, a.u.) (n=5). t-tests determined statistical significance. *p<0.05, **p<0.01.

Article Snippet: Sections were incubated overnight at 4°C with the following primary antibody anti-CD68 (1:200, Bio-Rad, MCA17576A), CD31 (1:250, Santa Cruz, sc-376764), MTTP (1:150, Thermo Fischer, PA5-76049) and CEPT1 (1:250, Bioss, bs-12284R-Cy3).

Techniques: Staining, Immunofluorescence

Journal: bioRxiv

Article Title: MTARC1 Regulates Lipid Droplet Degradation via Phospholipid Remodeling in Metabolic Fatty Liver Disease

doi: 10.1101/2025.07.17.665462

Figure Lengend Snippet: MTARC1 deficiency remodels hepatic glycerophospholipid homeostasis. (A) The workflow of proteomics and transcriptomics joint analysis. Briefly, eight livers from each group of mice challenged with CDAHFD for seven weeks were subjected to RNA-seq (n = 8/group). Two livers from the same group were pooled and subjected to TMT-based proteomics (n = 4/group). The hits with |Log 2 FC| > 0.3, P < 0.05 were defined as differentially expressed hits (DEHs). (B-C) Top four terms from gene ontology annotation for downregulated (B) and upregulated (C) hits (BP, biological process, CC, cellular component, MF, molecular function). (D) Heatmap presenting the detailed expression profile of the term “lipid metabolic process” from Panel C. (E) The simplified pathways through which PC and PE are synthesized. (F) RPKM level of Pemt and Cept1 mined from the transcriptomic data (n = 8/group). (G) Relative protein level of PEMT and CEPT1 mined from the proteomic data (n = 4/group). (H-I) qPCR and immunoblotting assays for livers from control and MKO mice challenged with CDAHFD for eight weeks. (J) Livers from control and MLKO mice challenged with CDAHFD for seven weeks were subjected to lipidomics analysis (n = 9/group). The relative total level of each subtype identified was presented in the heatmap. Data were expressed as mean ± SEM and analyzed by Student’s t-test. * p < 0.05, ** p < 0.01.

Article Snippet: Primary and secondary antibodies used in this study are listed below: MTARC1 (Aviva, ARP49748_P050), HSP60 (Proteintech, 15282-1-AP), COL1A1 (Zenbio, 501352), CEPT1 (Proteintech, 20496-1-AP), PEMT (Invitrogen, PA5-42383), Tubulin (Beyotime, AF0001), LAL (Abcam, ab154356), ATGL (Proteintech, 55190-1-AP), FAM134B (Proteintech, 21537-1-AP), Tom20 (Proteintech, 11802-1-AP), LAMP2 (Proteintech, 11802-1-AP), PMP70 (Abcam, ab85550), and ADRP (Zenbio, R381796).

Techniques: RNA Sequencing, Expressing, Synthesized, Western Blot, Control

Journal: bioRxiv

Article Title: MTARC1 Regulates Lipid Droplet Degradation via Phospholipid Remodeling in Metabolic Fatty Liver Disease

doi: 10.1101/2025.07.17.665462

Figure Lengend Snippet: Inhibition of glycerophospholipids biosynthesis reverses the hepatoprotective effect of MTARC1 deficiency. (A-C) Eight weeks old LSL Cas9 mice were administrated with the virus as indicated and subjected to CDAHFD challenge for 23 days (n = 9-10/group). The viral dosages (vg/mouse) are: 2e 10 for Pemt shRNA virus, 3e 10 for Cept1 shRNA, and 1e 11 for control virus. To ensure the equal amount of virus used for each mouse, control virus was used as the filler. (A) Animal diet challenge experiment workflow. (B) Growth curve. (C) Serum ALT activity. (D-M) Eight weeks old control and Mtarc1 knockout mice were administrated with virus as indicated and subjected to CDAHFD challenge for eight weeks (n = 9-10/group). (D) Animal experimental protocol. (E) Growth curve. (F) Liver body weight ratio (%). (G) Hepatic TG content. (H) Representative images of H&E staining for liver section (Scale bar, 50 μm). (I) Serum ALT activity. (J) Serum AST activity. (K-M) qPCR assay for Hepatic Col1a1 (K), Pemt (L), and Cept1 (M). Data were expressed as mean ± SEM and analyzed by Student’s t-test. * p < 0.05, ** p < 0.01.

Article Snippet: Primary and secondary antibodies used in this study are listed below: MTARC1 (Aviva, ARP49748_P050), HSP60 (Proteintech, 15282-1-AP), COL1A1 (Zenbio, 501352), CEPT1 (Proteintech, 20496-1-AP), PEMT (Invitrogen, PA5-42383), Tubulin (Beyotime, AF0001), LAL (Abcam, ab154356), ATGL (Proteintech, 55190-1-AP), FAM134B (Proteintech, 21537-1-AP), Tom20 (Proteintech, 11802-1-AP), LAMP2 (Proteintech, 11802-1-AP), PMP70 (Abcam, ab85550), and ADRP (Zenbio, R381796).

Techniques: Inhibition, Virus, shRNA, Control, Activity Assay, Knock-Out, Staining

Journal: Journal of Pharmaceutical Analysis

Article Title: Disorder of phospholipid metabolism in the renal cortex and medulla contributes to acute tubular necrosis in mice after cantharidin exposure using integrative lipidomics and spatial metabolomics

doi: 10.1016/j.jpha.2025.101210

Figure Lengend Snippet: The research strategy in cantharidin (CTD)-induced acute kidney injury (AKI) via lipidomics and spatial metabolomics. LC-MS/MS: liquid chromatography-tandem mass spectrometry; MSI: mass spectrometry imaging; UMAP: Uniform Manifold Approximation and Projection; PA: phosphatidic acid; PC: phosphatidylcholine; PE: phosphatidylethanolamine; PS: phosphatidylserine; LysoPC: lysophosphatidylcholine; SL: sphingolipid; SM: sphingomyelin; Cer: ceramide; CL: cardiolipin; DG: diacylglycerol; LPCAT: lysophosphatidylcholine acyltransferase; CEPT1: choline phosphotransferase 1.

Article Snippet: The blots were incubated with lysophosphatidylcholine acyltransferase 1 (LPCAT1) (1:1,000, Proteintech, Wuhan, China) and choline phosphotransferase 1 (CEPT1) (1:1,000, Proteintech, Wuhan, China), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:2,000, Proteintech, Wuhan, China) overnight at 4 °C.

Techniques: Liquid Chromatography with Mass Spectroscopy, Liquid Chromatography, Mass Spectrometry, Imaging

Journal: Journal of Pharmaceutical Analysis

Article Title: Disorder of phospholipid metabolism in the renal cortex and medulla contributes to acute tubular necrosis in mice after cantharidin exposure using integrative lipidomics and spatial metabolomics

doi: 10.1016/j.jpha.2025.101210

Figure Lengend Snippet: Integration analysis of lipidomics and spatial metabolomics in mouse kidney after cantharidin (CTD) exposure. (A) Venn diagram of common differential lipid metabolites. (B) Fold change (FC) value of common differential lipid metabolites. (C) Venn diagram of common differential lipid pathways. (D) Venn diagram of common of up- and down-regulated lipids in glycerophospholipid (GP) and sphingolipid (SL) metabolism. (E) A total of raw peak area of lipids in GP and SL metabolism. (F) Molecular docking results between CTD and lysophosphatidylcholine acyltransferase 1 (LPCAT1) and choline phosphotransferase 1 (CEPT1). UPLC-MS/MS: ultra-high performance liquid chromatography-tandem mass spectrometry; MALDI-MSI: matrix-assisted laser desorption/ionization-mass spectrometry imaging; PC: phosphatidylcholine; SM: sphingomyelin; PA: phosphatidic acid; PE: phosphatidyl ethanolamine; PS: phosphatidylserine; LysoPC: lysophosphatidylcholine.

Article Snippet: The blots were incubated with lysophosphatidylcholine acyltransferase 1 (LPCAT1) (1:1,000, Proteintech, Wuhan, China) and choline phosphotransferase 1 (CEPT1) (1:1,000, Proteintech, Wuhan, China), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:2,000, Proteintech, Wuhan, China) overnight at 4 °C.

Techniques: Tandem Mass Spectroscopy, High Performance Liquid Chromatography, Mass Spectrometry, Imaging

Journal: Journal of Pharmaceutical Analysis

Article Title: Disorder of phospholipid metabolism in the renal cortex and medulla contributes to acute tubular necrosis in mice after cantharidin exposure using integrative lipidomics and spatial metabolomics

doi: 10.1016/j.jpha.2025.101210

Figure Lengend Snippet: Spatial lipid metabolic mechanism in cantharidin (CTD)-induced kidney injury. The left part displays the lipids spatial distribution in kidney, and the right part displays the lipid metabolic mechanism. PA: phosphatidic acid; PC: phosphatidylcholine; PE: phosphatidylethanolamine; PS: phosphatidylserine; LysoPC: lysophosphatidylcholine; SL: sphingolipid; SM: sphingomyelin; Cer: ceramide; CL: cardiolipin; DG: diacylglycerol; LPCAT: lysophosphatidylcholine acyltransferase; CEPT1: choline phosphotransferase 1.

Article Snippet: The blots were incubated with lysophosphatidylcholine acyltransferase 1 (LPCAT1) (1:1,000, Proteintech, Wuhan, China) and choline phosphotransferase 1 (CEPT1) (1:1,000, Proteintech, Wuhan, China), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:2,000, Proteintech, Wuhan, China) overnight at 4 °C.

Techniques:

Journal: Journal of Pharmaceutical Analysis

Article Title: Disorder of phospholipid metabolism in the renal cortex and medulla contributes to acute tubular necrosis in mice after cantharidin exposure using integrative lipidomics and spatial metabolomics

doi: 10.1016/j.jpha.2025.101210

Figure Lengend Snippet: Accumulated lysophosphatidylcholine (LysoPC) (16:0/0:0) aggravated cantharidin (CTD)-induced acute tubular necrosis (ATN) in human kidney-2 (HK-2) cells. (A) The structure of LysoPC (16:0/0:0). (B) The LysoPC (16:0/0:0) content of the kidney using liquid chromatography-mass spectrometry (LC-MS) detection ( n = 6, mean ± standard error of the mean (SEM); ∗ P < 0.05 vs. the control group). (C) Cell viability detection after LysoPC (16:0/0:0) exposure in 12 h in HK-2 cells. (D) Cell viability detection after LysoPC (16:0/0:0) and CTD exposure in 12 h in HK-2 cells. (E, F) Molecular docking results between LysoPC (16:0/0:0) and choline phosphotransferase 1 (CEPT1) (E) and lysophosphatidylcholine acyltransferase 1 (LPCAT1) (F). (G, H) Relative protein expression level of LPCAT1 and CEPT1 in mouse kidney (G) and in HK-2 cells (H) after CTD intervention. n = 3, mean ± standard deviation (SD); ∗ P < 0.05, ∗∗ P < 0.01 vs. the control group.

Article Snippet: The blots were incubated with lysophosphatidylcholine acyltransferase 1 (LPCAT1) (1:1,000, Proteintech, Wuhan, China) and choline phosphotransferase 1 (CEPT1) (1:1,000, Proteintech, Wuhan, China), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:2,000, Proteintech, Wuhan, China) overnight at 4 °C.

Techniques: Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Control, Expressing, Standard Deviation