Journal: Life Science Alliance

Article Title: Inhibition of microRNA-33b in humanized mice ameliorates nonalcoholic steatohepatitis

doi: 10.26508/lsa.202301902

Figure Lengend Snippet: (A) Scheme of the HFD feeding protocol. (B) Representative microscopic images of HE staining of the liver of miR-33b −/− and miR-33b +/+ mice fed the HFD. Scale bars: 300 μm (upper). (C) Total cholesterol and triglyceride levels in the livers of miR-33b −/− and miR-33b +/+ mice fed the HFD. n = 4 mice per group; ** P < 0.01, unpaired t test. (D) Relative expression levels of miR-33 target genes in the livers of miR-33b −/− and miR-33b +/+ mice fed the HFD. n = 4–5 mice per group; ** P < 0.01, unpaired t test. (E) Western blotting analysis of ABCA1, CPT1A, and CROT expression in the livers of miR-33b −/− and miR-33b +/+ mice fed the HFD. GAPDH was used as a loading control. n = 5 mice per group. (F) Densitometry of hepatic ABCA1, CPT1A, and CROT. n = 5 mice per group; * P < 0.05 and ** P < 0.01, unpaired t test. (G) Relative expression levels of inflammatory genes and fibrosis-related genes in the livers of miR-33b −/− and miR-33b +/+ mice fed the HFD. n = 5–8 mice per group; ** P < 0.01, unpaired t test. (H) Western blotting analysis of COL1A1 expression in the livers of miR-33b −/− and miR-33b +/+ mice fed the HFD. GAPDH was used as a loading control. n = 5 mice per group. (I) Densitometry of hepatic COL1A1. n = 5 mice per group; * P < 0.05, unpaired t test. (J) Representative microscopic images of Masson’s trichrome staining of the livers of miR-33b −/− and miR-33b +/+ mice fed the HFD. Scale bars: 100 μm.

Article Snippet: The following antibodies were used: an anti-ABCA1 antibody (NB400-105; Novus Biologicals), anti-GAPDH antibody (#2118S; Cell Signaling Technology), anti-β-actin antibody (AC-15; A5441; Sigma-Aldrich), anti-COL1A1 antibody (ab34710; Abcam), anti-CROT antibody (ab103448; Abcam), anti-CPT1A antibody (ab128568; Abcam), anti-α-smooth muscle actin (SMA) antibody (1A4; C6198; Sigma-Aldrich), anti-F4/80 antibody (BM8; 123101; BioLegend), and anti-myeloperoxidase (MPO) antibody (C-3; sc-390109; Santa Cruz Biotechnology).

Techniques: Staining, Expressing, Western Blot, Control

Journal: Life Science Alliance

Article Title: Inhibition of microRNA-33b in humanized mice ameliorates nonalcoholic steatohepatitis

doi: 10.26508/lsa.202301902

Figure Lengend Snippet: (A) Body weight of miR-33b fl/fl KI mice and Ayu1 -Cre/miR-33b fl/fl KI mice at 8 wk of age. n = 5 mice per group. (B) Relative expression levels of miR-33b in the livers, epididymal white adipose tissues, hearts, and gastrocnemius muscles of miR-33b fl/fl KI mice and Ayu1 -Cre/miR-33b fl/fl KI mice. n = 4 mice per group; ** P < 0.01, unpaired t test. (C) Relative expression levels of miR-33a in the livers, epididymal white adipose tissues, hearts, and gastrocnemius muscles of miR-33b fl/fl KI mice and Ayu1 -Cre/miR-33b fl/fl KI mice. n = 4 mice per group. (D) Relative expression levels of Srebf1 , Srebf2 , Abca1 , Cpt1a , and Crot in the livers of miR-33b fl/fl KI mice and Ayu1 -Cre/miR-33b fl/fl KI mice. n = 4 mice per group; * P < 0.05, unpaired t test.

Article Snippet: The following antibodies were used: an anti-ABCA1 antibody (NB400-105; Novus Biologicals), anti-GAPDH antibody (#2118S; Cell Signaling Technology), anti-β-actin antibody (AC-15; A5441; Sigma-Aldrich), anti-COL1A1 antibody (ab34710; Abcam), anti-CROT antibody (ab103448; Abcam), anti-CPT1A antibody (ab128568; Abcam), anti-α-smooth muscle actin (SMA) antibody (1A4; C6198; Sigma-Aldrich), anti-F4/80 antibody (BM8; 123101; BioLegend), and anti-myeloperoxidase (MPO) antibody (C-3; sc-390109; Santa Cruz Biotechnology).

Techniques: Expressing, Muscles

Journal: Life Science Alliance

Article Title: Inhibition of microRNA-33b in humanized mice ameliorates nonalcoholic steatohepatitis

doi: 10.26508/lsa.202301902

Figure Lengend Snippet: (A) Representative microscopic images of HE (upper) and Picro-Sirius red (lower) staining of the livers of miR-33b fl/fl KI and Alb -Cre/miR-33b fl/fl KI mice fed the HFD. Scale bars: 200 μm. (B) Relative expression levels of Abca1 and Cpt1a in the livers of miR-33b fl/fl KI and Alb -Cre/miR-33b fl/fl KI mice fed the HFD. n = 10–11 mice per group; *** P < 0.001, unpaired t test. (C) Western blotting analysis of ABCA1 and CPT1A expression in the livers of miR-33b fl/fl KI and Alb -Cre/miR-33b fl/fl KI mice fed the HFD. GAPDH was used as a loading control. n = 5 mice per group. (D) Relative expression levels of Col1a1 , Acta2 , and Tnf in the livers of miR-33b fl/fl KI and Alb -Cre/miR-33b fl/fl KI mice fed the HFD. n = 9–10 mice per group; * P < 0.05, unpaired t test. (E) Quantification of the fibrosis area in the liver sections of miR-33b fl/fl KI and Alb -Cre/miR-33b fl/fl KI mice fed NC or the HFD according to Picro-Sirius red staining. n = 6–7 mice per group; *** P < 0.001, one-way ANOVA with Tukey’s post hoc test. (F) Total cholesterol content in the livers of miR-33b fl/fl KI and Alb -Cre/miR-33b fl/fl KI mice fed NC or the HFD. n = 6–7 mice per group; * P < 0.05 and *** P < 0.001, one-way ANOVA with Tukey’s post hoc test. (G) Triglyceride content in the livers of miR-33b fl/fl KI and Alb -Cre/miR-33b fl/fl KI mice fed NC or the HFD. n = 6–7 mice per group; *** P < 0.001, one-way ANOVA with Tukey’s post hoc test.

Article Snippet: The following antibodies were used: an anti-ABCA1 antibody (NB400-105; Novus Biologicals), anti-GAPDH antibody (#2118S; Cell Signaling Technology), anti-β-actin antibody (AC-15; A5441; Sigma-Aldrich), anti-COL1A1 antibody (ab34710; Abcam), anti-CROT antibody (ab103448; Abcam), anti-CPT1A antibody (ab128568; Abcam), anti-α-smooth muscle actin (SMA) antibody (1A4; C6198; Sigma-Aldrich), anti-F4/80 antibody (BM8; 123101; BioLegend), and anti-myeloperoxidase (MPO) antibody (C-3; sc-390109; Santa Cruz Biotechnology).

Techniques: Staining, Expressing, Western Blot, Control

Journal: Life Science Alliance

Article Title: Inhibition of microRNA-33b in humanized mice ameliorates nonalcoholic steatohepatitis

doi: 10.26508/lsa.202301902

Figure Lengend Snippet: (A) Relative expression levels of miR-33b and miR-33a in the peritoneal macrophages of miR-33b fl/fl KI mice and LysM -Cre/miR-33b fl/fl KI mice. n = 4 samples per group; *** P < 0.01, unpaired t test. (B) Body weight change of miR-33b fl/fl KI mice and LysM -Cre/miR-33b fl/fl KI mice under HFD feeding. (C) Body weight at the time of euthanasia of miR-33b fl/fl KI mice and LysM -Cre/miR-33b fl/fl KI mice fed the HFD. n = 9–14 mice per group. (D) Relative expression levels of Abca1 , Cpt1a , Crot , Tnf , Il6 , Col1a1 , and Acta2 in the livers of miR-33b fl/fl KI mice and LysM -Cre/miR-33b fl/fl KI mice fed the HFD. n = 7 mice per group. (E) Representative microscopic images of HE staining of the liver of miR-33b fl/fl KI mice and LysM -Cre/miR-33b fl/fl KI mice fed the HFD. Scale bars: 200 μm.

Article Snippet: The following antibodies were used: an anti-ABCA1 antibody (NB400-105; Novus Biologicals), anti-GAPDH antibody (#2118S; Cell Signaling Technology), anti-β-actin antibody (AC-15; A5441; Sigma-Aldrich), anti-COL1A1 antibody (ab34710; Abcam), anti-CROT antibody (ab103448; Abcam), anti-CPT1A antibody (ab128568; Abcam), anti-α-smooth muscle actin (SMA) antibody (1A4; C6198; Sigma-Aldrich), anti-F4/80 antibody (BM8; 123101; BioLegend), and anti-myeloperoxidase (MPO) antibody (C-3; sc-390109; Santa Cruz Biotechnology).

Techniques: Expressing, Staining

Journal: Life Science Alliance

Article Title: Inhibition of microRNA-33b in humanized mice ameliorates nonalcoholic steatohepatitis

doi: 10.26508/lsa.202301902

Figure Lengend Snippet: (A) Relative expression levels of Abca1 , Cpt1a , and Crot in the livers of the indicated groups. n = 6–10 samples per group; * P < 0.05 and *** P < 0.001, one-way ANOVA with Tukey’s post hoc test. (B) Western blotting analysis of ABCA1, CPT1A, and CROT expression in the livers of the indicated groups. GAPDH was used as a loading control. n = 5 samples per group. (C) Densitometry of hepatic ABCA1, CPT1A, and CROT. n = 5 mice per group; * P < 0.05 and ** P < 0.01, unpaired t test. (D) Total cholesterol, free cholesterol, and triglyceride levels in the livers of the indicated groups. n = 4–7 samples per group; ** P < 0.01 and *** P < 0.001, one-way ANOVA with Tukey’s post hoc test. (E) Representative images of cholesterol crystals with and without a polarizing filter in the liver sections of the indicated groups. (F) Quantification of cholesterol crystal in the liver sections. n = 4–6 samples per group; ** P < 0.01 and *** P < 0.001, one-way ANOVA with Tukey’s post hoc test.

Article Snippet: The following antibodies were used: an anti-ABCA1 antibody (NB400-105; Novus Biologicals), anti-GAPDH antibody (#2118S; Cell Signaling Technology), anti-β-actin antibody (AC-15; A5441; Sigma-Aldrich), anti-COL1A1 antibody (ab34710; Abcam), anti-CROT antibody (ab103448; Abcam), anti-CPT1A antibody (ab128568; Abcam), anti-α-smooth muscle actin (SMA) antibody (1A4; C6198; Sigma-Aldrich), anti-F4/80 antibody (BM8; 123101; BioLegend), and anti-myeloperoxidase (MPO) antibody (C-3; sc-390109; Santa Cruz Biotechnology).

Techniques: Expressing, Western Blot, Control

Journal: Life Science Alliance

Article Title: Inhibition of microRNA-33b in humanized mice ameliorates nonalcoholic steatohepatitis

doi: 10.26508/lsa.202301902

Figure Lengend Snippet: (A) Relative expression levels of miR-33b and miR-33a in LI90 cells treated with a negative control or anti-miR-33b. n = 3 cells per group; ** P < 0.01, unpaired t test. (B) Relative expression levels of miR-33b in LI90 cells treated or not treated with TGF-β. n = 3 cells per group; *** P < 0.001, unpaired t test. (C) Relative expression levels of ABCA1 , CPT1A , and CROT in LI90 cells treated with a negative control or anti-miR-33b. n = 4 cells per group. (D) Western blotting analysis of ABCA1, CPT1A, and CROT expression in LI90 cells treated with a negative control or anti-miR-33b. n = 4 cells per group. (E) Densitometry of ABCA1, CPT1A, and CROT in LI90 cells treated with a negative control or anti-miR-33b. n = 4 cells per group; ** P < 0.01 and *** P < 0.001, unpaired t test. (F) Relative expression levels of COL1A1 , ACTA2 , and POSTN in LI90 cells treated with a negative control or anti-miR-33b. n = 4 cells per group.

Article Snippet: The following antibodies were used: an anti-ABCA1 antibody (NB400-105; Novus Biologicals), anti-GAPDH antibody (#2118S; Cell Signaling Technology), anti-β-actin antibody (AC-15; A5441; Sigma-Aldrich), anti-COL1A1 antibody (ab34710; Abcam), anti-CROT antibody (ab103448; Abcam), anti-CPT1A antibody (ab128568; Abcam), anti-α-smooth muscle actin (SMA) antibody (1A4; C6198; Sigma-Aldrich), anti-F4/80 antibody (BM8; 123101; BioLegend), and anti-myeloperoxidase (MPO) antibody (C-3; sc-390109; Santa Cruz Biotechnology).

Techniques: Expressing, Negative Control, Western Blot

Journal: Frontiers in Immunology

Article Title: Ex vivo expanded human regulatory T cells promote cholesterol efflux and PON1 expression in oxLDL-exposed macrophages via gap junction-mediated cAMP transfer

doi: 10.3389/fimmu.2025.1662925

Figure Lengend Snippet: Treg exp increase ABCA1 expression by transferring cAMP into M IL4 . (A) Changes in the expression of ABCA1 and ABCG1 transcripts in M IL4 co-cultured with either Teffs or Treg exp . Gene expression quantified by qRT-PCR comparing mRNA levels in M IL4 alone (RQ = 1, shown as dashed line) with the same cells co-cultured for 4h with either Teffs or Tregs. UBC was used as reference gene. (B) Intracellular concentration of cAMP in Tregs exp alone, M IL4 alone, or M IL4 co-cultured with Tregs exp for 4h. cAMP levels were normalized to total cellular protein content. Statistical analysis was performed only between M IL4 and M IL4 + Tregs exp , as the aim was to assess whether cAMP levels in M IL4 increase upon co-culture. The cAMP level in Tregs exp is shown as a reference to highlight the high concentration of this molecule in these cells but was not included in the statistical comparison due to the difference in cell type. (C) Changes in the expression of ABCA1 mRNA in M IL4 co-cultured with Treg exp (ratio 1:1) after 1h preincubation or not with GAP27 (300 µM). (D) Changes in the expression of ABCA1 mRNA in M IL4 co-cultured with Tregs (ratio 1:1) after 1h preincubation or not with PKA inhibitor H89 (5 µM). (E) Western blot analysis of ABCA1 protein level in cell lysates of M IL4 alone or M IL4 co-cultured (ratio 1:1) with either Treg exp or Teff for 4h. Data were plotted as ABCA1 protein intensity normalised to GAPDH protein intensity. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparison test. *p<0.05, **p<0.01.

Article Snippet: Equal amounts of total protein were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), transferred to polyvinylidene difluoride (PVDF) membrane and probed overnight at 4C with the respective antibodies: monoclonal anti-ABCA1 antibody (CellSignaling; Cat Number 96292), polyclonal anti-PON1 antibody (Proteintech; Cat Number 18155-1-AP), monoclonal anti-GAPDH antibody (CellSignaling; Cat Number 5174), recombinant monoclonal anti-ABCA1 (phosho-S2054) antibody (Abcam; Cat Number ab125064).

Techniques: Expressing, Transferring, Cell Culture, Gene Expression, Quantitative RT-PCR, Concentration Assay, Co-Culture Assay, Comparison, Western Blot

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Dysregulated cholesterol uptake and efflux of bone marrow-derived α-SMA + macrophages contribute to atherosclerotic plaque formation

doi: 10.1007/s00018-025-05655-3

Figure Lengend Snippet: α-SMA promotes the lipid uptake and inhibits lipid efflux in macrophages. A , B Flow cytometric analysis of binding and uptake of DiI-Ox-LDL in vector- or Acta2 hi RAW264.7 cells A , and BMDMs from Acta2 f/f or Acta2 MKO mice B . The quantification results are shown on the right. n = 3. * P < 0.05, *** P < 0.001, **** P < 0.0001 by unpaired Student’s t-test. C , D SR-A, ABCA1, ABCG1 and α-SMA protein levels in vector- and Acta2 hi RAW264.7 cells C , and BMDMs from Acta2 Flox or Acta2 MKO mice D . n = 3. * P < 0.05 by unpaired Student’s t-test. E–F Co-staining and quantification of SR-A + MOMA-2 + E or ABCA1 + MOMA-2 + F cells in aortic roots from Acta2 f/f or Acta2 MKO mice treated with AAV -PCSK9 DY followed by a 12-week HFD. n = 7. * P < 0.05 by unpaired Student’s t-test. G–H Flow cytometric analysis of binding G and uptake H of DiI-Ox-LDL in Acta2 hi RAW264.7 cells treated with an SR-A blocking antibody (20 μg/mL) or IgG control for 2 h were detected by flow cytometry. n = 3. * P < 0.05 by unpaired Student’s t-test. AAV, adeno-associated virus; HFD, high-fat diet

Article Snippet: Primary antibodies used included mouse anti-mouse α-SMA (BM0002, Boster, USA), goat anti-mouse SR-A (af1797, R&D, USA), rabbit anti-mouse ABCA1 (PB0490, Boster, USA), human anti-mouse ABCG1 (bs-23382R, Bioss, China), rabbit anti-human CD36 (ab133625, Abcam, USA), mouse GAPDH (AF0006, Beyotime, China), rabbit anti-mouse p-Smad3 (AF1759, Beyotime, China), and rabbit anti-mammalian Smad3 (sc-101154, SANTA CRUZ, USA).

Techniques: Binding Assay, Plasmid Preparation, Staining, Blocking Assay, Control, Flow Cytometry, Virus

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Dysregulated cholesterol uptake and efflux of bone marrow-derived α-SMA + macrophages contribute to atherosclerotic plaque formation

doi: 10.1007/s00018-025-05655-3

Figure Lengend Snippet: AKT pathway is involved in α-SMA-induced lipid accumulation. A Abca1 mRNA levels in vector or Acta2 hi RAW264.7 cells treated with 50 μg/ml Ox-LDL for 0, 6, 12, or 24 h. n = 3. * P < 0.05 by unpaired Student’s t-test. B Top 15 KEGG pathways of genes downregulated in Acta2 hi RAW264.7 cells incubated with Ox-LDL for 6 h. C Protein levels of p-AKT, AKT, p-ERK, ERK, p-STAT3, STAT3, CD36, SR-A, ABCA1 and α-SMA treated with 50 μg/ml Ox-LDL for 6 h. n = 3. * P < 0.05 by one-way ANOVA. D – F Flow cytometric analysis of binding ( D and E ) and uptake ( D and F ) of DiI-Ox-LDL in vector or Acta2 hi RAW264.7 cells incubated with or without SC79 (2 μg/mL). n = 3. * P < 0.05, ** P < 0.01, **** P < 0.0001 by one-way ANOVA. G – I Protein levels of CD36, SR-A, ABCA1 and α-SMA in Acta2 hi or vector RAW264.7 cells incubated with or without SC79 (2 μg/mL) and treated with 50 μg/ml Ox-LDL for 2 h. n = 3. * P < 0.05, ** P < 0.01, **** P < 0.0001 by one-way ANOVA

Article Snippet: Primary antibodies used included mouse anti-mouse α-SMA (BM0002, Boster, USA), goat anti-mouse SR-A (af1797, R&D, USA), rabbit anti-mouse ABCA1 (PB0490, Boster, USA), human anti-mouse ABCG1 (bs-23382R, Bioss, China), rabbit anti-human CD36 (ab133625, Abcam, USA), mouse GAPDH (AF0006, Beyotime, China), rabbit anti-mouse p-Smad3 (AF1759, Beyotime, China), and rabbit anti-mammalian Smad3 (sc-101154, SANTA CRUZ, USA).

Techniques: Plasmid Preparation, Incubation, Binding Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Dysregulated cholesterol uptake and efflux of bone marrow-derived α-SMA + macrophages contribute to atherosclerotic plaque formation

doi: 10.1007/s00018-025-05655-3

Figure Lengend Snippet: Schematic diagram illustrating the role of bone marrow-derived α-SMA + macrophages in atherosclerotic plaque formation. Macrophage-expressed α-SMA enhances SR-A expression via the AKT signaling pathway, leading to increased lipid binding and uptake. Concurrently, α-SMA suppresses ABCA1 expression, thereby reducing lipid efflux. This imbalance promotes lipid accumulation and contributes to atherosclerotic plaque formation

Article Snippet: Primary antibodies used included mouse anti-mouse α-SMA (BM0002, Boster, USA), goat anti-mouse SR-A (af1797, R&D, USA), rabbit anti-mouse ABCA1 (PB0490, Boster, USA), human anti-mouse ABCG1 (bs-23382R, Bioss, China), rabbit anti-human CD36 (ab133625, Abcam, USA), mouse GAPDH (AF0006, Beyotime, China), rabbit anti-mouse p-Smad3 (AF1759, Beyotime, China), and rabbit anti-mammalian Smad3 (sc-101154, SANTA CRUZ, USA).

Techniques: Derivative Assay, Expressing, Binding Assay

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: ATP-binding cassette transporter 1 participates in LDL oxidation by artery wall cells.

doi: 10.1161/01.atv.0000035700.82829.2a

Figure Lengend Snippet: Figure 1. Glyburide and DIDS, inhibitors of ABCA1 function, impair cell-mediated modification of LDL in human artery wall cocultures. Artery wall cocultures were treated overnight with either glyburide (100 mol/L) or DIDS (100 mol/L) in medium 199 containing 10% lipoprotein-deficient serum (LPDS). LDL (250 g/mL) was added to the cocultures, and 18 hours later, the supernatants were removed and assayed for Auerbach lipid hydroperoxide equivalents30 (left) and monocyte chemotactic activity (right). Similar results were obtained in 3 independent experiments. *P0.05.

Article Snippet: Proteins were electroblotted onto Hybond ECL Nitrocellulose membranes (Amersham) and incubated for 3 hours with rabbit anti-human ABCA1 antiserum (Novus Biologicals) at 1:100 dilution.

Techniques: Activity Assay

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: ATP-binding cassette transporter 1 participates in LDL oxidation by artery wall cells.

doi: 10.1161/01.atv.0000035700.82829.2a

Figure Lengend Snippet: Figure 4. Antisense oligonucleotides to ABCA1 inhibit cell- mediated modification of LDL in22-R-OHC–treated human artery wall cocultures. Artery wall cocultures were transfected with either antisense or sense phosphorothioate oligonucleotides (100 nmol/L) to human ABCA1, as described in Methods. The cells were further treated with 22-R-OHC (25 mol/L) in the presence or absence of LDL (250 g/mL). After the treatments, cell lysates were prepared and analyzed by Western blotting for ABCA1 protein expression (A), and cell culture supernatants were analyzed for monocyte chemotactic activity (B). *P0.05.

Article Snippet: Proteins were electroblotted onto Hybond ECL Nitrocellulose membranes (Amersham) and incubated for 3 hours with rabbit anti-human ABCA1 antiserum (Novus Biologicals) at 1:100 dilution.

Techniques: Transfection, Western Blot, Expressing, Cell Culture, Activity Assay

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: ATP-binding cassette transporter 1 participates in LDL oxidation by artery wall cells.

doi: 10.1161/01.atv.0000035700.82829.2a

Figure Lengend Snippet: Figure 2. 22-R-OHC induces ABCA1 expression and enhances cell-mediated modification of LDL in human artery wall cocul- tures. A and B, Confluent cocultures (HASMCs and HAECs) were either untreated () or treated () with 22-R-OHC (25 mol/L). Eighteen hours later, the cells were lysed, and 10 g total RNA (A) and 60 g total protein (B) from each condition were subjected to Northern and Western analyses for ABCA1, respectively. C, Artery wall cocultures were either left untreated or were treated with 22-R-OHC (25 mol/L) for 18 hours. Freshly isolated LDL (250 g/mL) was added to cells, and 6 hours later, supernatants were removed and assayed for Auer- bach lipid hydroperoxide equivalents30 (left) and monocyte che- motactic activity (right) as described in Methods. Each experi- mental condition was performed in triplicate wells, and data are expressed as meanSD. The results were similar in 3 separate experiments. *P0.05.

Article Snippet: Proteins were electroblotted onto Hybond ECL Nitrocellulose membranes (Amersham) and incubated for 3 hours with rabbit anti-human ABCA1 antiserum (Novus Biologicals) at 1:100 dilution.

Techniques: Expressing, Northern Blot, Western Blot, Isolation, Activity Assay

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: ATP-binding cassette transporter 1 participates in LDL oxidation by artery wall cells.

doi: 10.1161/01.atv.0000035700.82829.2a

Figure Lengend Snippet: Figure 3. Antisense oligonucleotides to ABCA1 inhibit artery wall cell–mediated modification of LDL. Artery wall cocultures were transfected with antisense phosphorothioate oligonucleo- tides (100 nmol/L) to human ABCA1. After transfections, control and transfected cells were either left untreated or were treated with LDL (250 g/mL) for an additional 6 hours. After the treat- ments, cell supernatants were collected and analyzed for Auer- bach lipid hydroperoxide equivalents30 (left) and monocyte che- motactic activity (right). The results were similar in 3 independent experiments. *P0.05.

Article Snippet: Proteins were electroblotted onto Hybond ECL Nitrocellulose membranes (Amersham) and incubated for 3 hours with rabbit anti-human ABCA1 antiserum (Novus Biologicals) at 1:100 dilution.

Techniques: Transfection, Control, Activity Assay

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: ATP-binding cassette transporter 1 participates in LDL oxidation by artery wall cells.

doi: 10.1161/01.atv.0000035700.82829.2a

Figure Lengend Snippet: Figure 6. ABCA1 and apoA-I are components of the mechanism that links reverse cholesterol transport and LDL oxidation. In the artery wall cells (HAECs and HASMCs), ABCA1 is (either directly or indirectly) required for the transport of cholesterol, phospho- lipids, and oxidized lipids. ApoA-I acquires cholesterol and phospholipids in an ABCA1-dependent mechanism and partici- pates in HDL synthesis.11 Lipoxygenase-derived hydroperoxides or other ROS are transferred across the cell membrane via an ABCA1-dependent mechanism to seed the extracellular LDL. ApoA-I removes oxidized lipids and ROS and renders artery wall cells unable to modify LDL.7,8 The diagram also assumes that the cholesterol content of the cells determines the cellular con- tent of oxygenated sterols, such as 22-R-OHC. 22-R-OHC increases cellular ROS in the presence of LDL by an as-yet- unidentified pathway and increases LDL oxidation. Thus, the reverse cholesterol transport pathway may be linked to LDL oxi- dation in at least 2 ways: (1) ABCA1 is required for reverse cho- lesterol transport and LDL oxidation. (2) Cellular cholesterol lev- els may determine the cellular levels of 22-R-OHC, which, in part, regulates cell-mediated LDL oxidation by an as-yet- unidentified pathway.

Article Snippet: Proteins were electroblotted onto Hybond ECL Nitrocellulose membranes (Amersham) and incubated for 3 hours with rabbit anti-human ABCA1 antiserum (Novus Biologicals) at 1:100 dilution.

Techniques: Derivative Assay, Membrane