Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Smooth Muscle Cells Contribute the Majority of Foam Cells in Apolipoprotein E-Deficient Mouse Atherosclerosis

doi: 10.1161/ATVBAHA.119.312434

Figure Lengend Snippet: (A) ABCA1 expression of CD45+ and CD45− foam cells from ApoE−/− mice fed a WD for 6 weeks (left panel) or 27-week-old male ApoE−/− mice fed a chow diet (right panel) determined as the median fluorescence of ABCA1 intensity by flow cytometry. For WD-fed mice, **P < 0.01 using two-way ANOVA, n=9 mice/group; there was no statistical difference between the two sexes. For chow-fed mice, **P < 0.01 using Mann–Whitney U test, n=5 mice/group. (B) Fold change of ABCA1 mRNA level in RAW 264.7 macrophages and MASMCs treated with 100 g/ml agLDL as compared to fatty acid-free albumin alone for 24 h. *P < 0.05, using Welch’s t test. (C) ABCA1 protein levels and cholesterol efflux from RAW 264.7 macrophages and MASMCs treated with 100 μg/mL agLDL for 48 hrs followed by incubation with 0.3 mM 8Br-cAMP ± 10 μg/mL (for efflux) for 24h. Cholesterol efflux was determined by LC-MS and normalized by total cell protein levels *P < 0.05, **P < 0.01 using two-tailed Mann–Whitney U test, N=5.

Article Snippet: BV421 rat anti-mouse CD45, V500 rat anti-mouse I-A/I-E (BD Pharmingen), and Alexa Fluor647 rat anti-mouse ABCA1 (AbD Serotec) antibodies were use according to manufacturer’s instructions.

Techniques: Expressing, Fluorescence, Flow Cytometry, MANN-WHITNEY, Incubation, Liquid Chromatography with Mass Spectroscopy, Two Tailed Test

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Smooth Muscle Cells Contribute the Majority of Foam Cells in Apolipoprotein E-Deficient Mouse Atherosclerosis

doi: 10.1161/ATVBAHA.119.312434

Figure Lengend Snippet: In humans, a thick diffuse intimal thickening (DIT) layer of SMCs is present in atherosclerosis-prone arteries beginning from birth8. At the onset of atherosclerosis lipoproteins that diffuse into the artery wall are trapped through a charge-charge interaction with SMC-secreted proteoglycans, primarily in the deep intima7. These trapped lipoproteins are modified and aggregated, becoming a substrate for uptake by the surrounding SMCs. At this stage monocyte/macrophages are located primarily in the subendothelial region away from the deposited lipids9. Over time both SMCs and macrophages take up modified lipoproteins to become foam cells independently and/or interactively. ABCA1 (red dots), the rate-limiting cholesterol efflux promoter, is robustly upregulated in macrophage foam cells, but less so in SMC foam cells. Increased ABCA1 expression facilitates cholesterol removal out of macrophage-derived foam cells, while impaired ABCA1 expression reduces cholesterol efflux from SMC foam cells, resulting in SMCs being the predominant lineage among total foam cells. In ApoE−/− mice, no DIT SMC layer is present. Atherogenic lipoproteins induce endothelial injury and monocyte binding and infiltration into the intima, where they mature into macrophages that take up the modified intimal lipids. Cytokines released by injured endothelial cells and monocytes/macrophages, including PDGF, TNF-α, and IL-1β induce SMC migration from the media into the intima. SMCs proliferate in the intima and take up modified lipoproteins to become foam cells. Similar to human lesions, the SMC foam cells exhibit reduced ABCA1 expression compared to macrophages. Over time the total contribution to foam cells shifts from macrophages to SMCs, due at least in part to impaired ABCA1 expression by the SMC foam cells.

Article Snippet: BV421 rat anti-mouse CD45, V500 rat anti-mouse I-A/I-E (BD Pharmingen), and Alexa Fluor647 rat anti-mouse ABCA1 (AbD Serotec) antibodies were use according to manufacturer’s instructions.

Techniques: Modification, Expressing, Derivative Assay, Binding Assay, Migration

Journal: Frontiers in Pharmacology

Article Title: Investigation of the Lipid-Lowering Mechanisms and Active Ingredients of Danhe Granule on Hyperlipidemia Based on Systems Pharmacology

doi: 10.3389/fphar.2020.00528

Figure Lengend Snippet: The detail information of three pathways mainly involved in lipid metabolism.

Article Snippet: Anti-SREBP-1c, anti-liver X receptor alpha (LXRα), anti-cholesterol 7α-hydroxylase (CYP7A1), and anti-ATP binding cassette subfamily A member 1 (ABCA1) were purchased from ImmunoWay Biotechnology Company (Newark, DE, USA).

Techniques: Control

Journal: Frontiers in Pharmacology

Article Title: Investigation of the Lipid-Lowering Mechanisms and Active Ingredients of Danhe Granule on Hyperlipidemia Based on Systems Pharmacology

doi: 10.3389/fphar.2020.00528

Figure Lengend Snippet: Effect of DHG on mRNA expression of genes in hamster liver by RT-PCR. Values are mean ± SD, n = 5. (A) SREBP-1c mRNA. (B) FAS mRNA. (C) SREBP-2 mRNA. (D) LDLR mRNA. (E) PPARα mRNA. (F) LXRα mRNA. (G) CYP7A1 mRNA. (H) ABCA1 mRNA. RT-PCR, real-time polymerase chain reaction; SREBP-1c, sterol regulatory element-binding protein-1c; FAS, fatty acid synthase; SREBP-2, sterol regulatory element binding protein 2; LDLR, low density lipoprotein receptor; PPARα, peroxisome proliferation-activated receptor alpha; LXRα, liver X receptor alpha; CYP7A1, cholesterol 7α-hydroxylase; ABCA1, ATP binding cassette subfamily A member 1. * p < 0.05 vs NC; ** p < 0.01 vs NC; # p < 0.05 vs HFD; ## p < 0.01 vs HFD.

Article Snippet: Anti-SREBP-1c, anti-liver X receptor alpha (LXRα), anti-cholesterol 7α-hydroxylase (CYP7A1), and anti-ATP binding cassette subfamily A member 1 (ABCA1) were purchased from ImmunoWay Biotechnology Company (Newark, DE, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Binding Assay

Journal: Frontiers in Pharmacology

Article Title: Investigation of the Lipid-Lowering Mechanisms and Active Ingredients of Danhe Granule on Hyperlipidemia Based on Systems Pharmacology

doi: 10.3389/fphar.2020.00528

Figure Lengend Snippet: Effect of DHG on protein expression of targets in hamster liver by Western blot. Values are mean ± SD, n = 3. (A) SREBP-1c, FAS. (B) SREBP-2, LDLR. (C) PPARα, LXRα, CYP7A1. (D) ABCA1. * p < 0.05, ** p < 0.01 vs NC group; # p < 0.05, ## p < 0.01 vs HFD group. The expression levels were normalized by GAPDH.

Article Snippet: Anti-SREBP-1c, anti-liver X receptor alpha (LXRα), anti-cholesterol 7α-hydroxylase (CYP7A1), and anti-ATP binding cassette subfamily A member 1 (ABCA1) were purchased from ImmunoWay Biotechnology Company (Newark, DE, USA).

Techniques: Expressing, Western Blot

Journal: International Journal of Oral Science

Article Title: Antisense oligonucleotide targeting TARDBP-EGFR splicing axis inhibits progression of oral squamous cell carcinoma through ABCA1-regulated cholesterol efflux

doi: 10.1038/s41368-025-00402-7

Figure Lengend Snippet: EGFR isoforms differentially inhibit cell proliferation by facilitating ABCA1 activation. a Western blotting analysis of the effect of EGFR isoform overexpression on the phosphorylation of EGFR. b , c Immunohistochemistry (IHC) staining of pEGFR and ABCA1. Scale bars, 100 μm (left), 25 μm (right). d The protein expression of ABCA1 in HN6 and CAL27 cells overexpressing EGFR-001 or EGFR-004 was detected by western blotting. e Co-immunoprecipitation analysis of the interaction between ABCA1 and EGFR in HN6 and CAL27 cells transfected with control, EGFR-001 , or EGFR-004 plasmids

Article Snippet: The lysates were then incubated overnight at 4 °C with either an anti-ABCA1 antibody (DF8233, Affinity Biosciences) or IgG (30000-0-AP, Proteintech) with constant rotation.

Techniques: Activation Assay, Western Blot, Over Expression, Phospho-proteomics, Immunohistochemistry, Expressing, Immunoprecipitation, Transfection, Control

Journal: International Journal of Oral Science

Article Title: Antisense oligonucleotide targeting TARDBP-EGFR splicing axis inhibits progression of oral squamous cell carcinoma through ABCA1-regulated cholesterol efflux

doi: 10.1038/s41368-025-00402-7

Figure Lengend Snippet: Ubiquitination levels of different EGFR isoforms. a Western blotting showing the ABCA1 protein in CAL27 and HN6 cells transfected with control, EGFR-001 , or EGFR-004 plasmids and treated with CHX (20 μg/mL) for the indicated times. b Western blotting of ABCA1 in HN6 and CAL27 cells transfected with EGFR-001 , EGFR-004 or control vectors, followed by treatment with 20 μmol/L MG132 for 6 h. c Ubiquitination level of ABCA1 protein in HN6 and CAL27 cells treated with MG132 and overexpressed with EGFR-001 , EGFR-004 were detected by western blotting

Article Snippet: The lysates were then incubated overnight at 4 °C with either an anti-ABCA1 antibody (DF8233, Affinity Biosciences) or IgG (30000-0-AP, Proteintech) with constant rotation.

Techniques: Ubiquitin Proteomics, Western Blot, Transfection, Control

Journal: International Journal of Oral Science

Article Title: Antisense oligonucleotide targeting TARDBP-EGFR splicing axis inhibits progression of oral squamous cell carcinoma through ABCA1-regulated cholesterol efflux

doi: 10.1038/s41368-025-00402-7

Figure Lengend Snippet: ABCA1 modulates membrane fluidity and Vimentin-mediated EMT in OSCC cells via cholesterol efflux. a Relative membrane cholesterol levels after overexpression of different EGFR isoforms. b Filipin staining was performed after ABCA1 was knocked down. c Total cholesterol contents after ABCA1 knockdown. d Membrane cholesterol levels were measured after ABCA1 knockdown. e Protein expression of Vimentin in HN6 and CAL27 cells treated with cholesterol and MβCD. f Effects of siABCA1 and cholesterol on the expression of Vimentin protein in HN6 and CAL27 cells. Statistical significance was assessed with Student’s t -test. * P < 0.05, ** P < 0.01, ** *P < 0.00 1

Article Snippet: The lysates were then incubated overnight at 4 °C with either an anti-ABCA1 antibody (DF8233, Affinity Biosciences) or IgG (30000-0-AP, Proteintech) with constant rotation.

Techniques: Membrane, Over Expression, Staining, Knockdown, Expressing

Journal: International Journal of Oral Science

Article Title: Antisense oligonucleotide targeting TARDBP-EGFR splicing axis inhibits progression of oral squamous cell carcinoma through ABCA1-regulated cholesterol efflux

doi: 10.1038/s41368-025-00402-7

Figure Lengend Snippet: Schematic illustration of the function and mechanism of the rs737540 in OSCC risk (By Figdraw). Compared with the rs737540-C allele, the risk rs737540-T allele promotes the inclusion of exon 4 of EGFR mediated by TARDBP to increase the expression of the full-length EGFR isoform ( EGFR-001 ) and decrease the expression level of the truncated EGFR isoform ( EGFR-004 ). Compared with the EGFR-004 subtype without exon 4, EGFR-001 was more able to inhibit ABCA1 ubiquitination and degradation by inhibiting EGFR phosphorylation, thus promoting OSCC cell proliferation, which provided functional evidence supporting our findings in a large population that rs737540-T contributes to increased OSCC risk. Meanwhile, ABCA1 can affect membrane fluidity by regulating membrane cholesterol levels. In addition, ASO targeting rs737540-T showed a significant tumor suppressor effect. The combination of ASO and Cetuximab shows broad prospects

Article Snippet: The lysates were then incubated overnight at 4 °C with either an anti-ABCA1 antibody (DF8233, Affinity Biosciences) or IgG (30000-0-AP, Proteintech) with constant rotation.

Techniques: Expressing, Ubiquitin Proteomics, Phospho-proteomics, Functional Assay, Membrane