mouse anti abca1  (Santa Cruz Biotechnology)

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: Fig. 4 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 Acta2hi RAW264.7 cells A, and BMDMs from Acta2f/f or Acta2MKO 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 Acta2hi RAW264.7 cells C, and BMDMs from Acta2Flox or Acta2MKO mice D. n = 3. *P < 0.05 by unpaired Student’s t-test. E–F Co-staining and quantification of SR-

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

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: Fig. 5 AKT pathway is involved in α-SMA-induced lipid accumu- lation. A Abca1 mRNA levels in vector or Acta2hi 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 Acta2hi 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 cyto-

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

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: Fig. 6 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: Acta Pharmaceutica Sinica. B

Article Title: PPAR α affects hepatic lipid homeostasis by perturbing necroptosis signals in the intestinal epithelium

doi: 10.1016/j.apsb.2024.08.021

Figure Lengend Snippet: Pparα deficiency in intestinal epithelium increases the translocation of gut-derived antigens into the liver. (A) Intestinal permeability assessment (FITC-dextran, 4 kD) in 8-week-old mice ( n = 10). (B) Relative mRNA levels of Zo-1 and Cldn8 in the ileum from 8-week-old mice ( n = 5). (C) The relative proportion of bacterial species in the cecum content by 16S rRNA gene sequencing ( n = 6). (D) Bugbase phenotypic prediction of gut microbiota in 8-week-old mice ( n = 6). (E) The mRNA and protein levels of PV1 in the ileum of 8-week-old mice ( n = 5). (F) Transmission electron microscopy images of the diaphragm (red star) in the capillaries from ileum sections in 24-week-old mice ( n = 3). (G) Representative images of fluorescence microscopy and transmission electron in 8-week-old mice treated with FITC-LPS (green) or Au-LPS ( n = 3–5). (H) Portal HDL-C levels in 8-week-old mice ( n = 10). (I) The protein levels of APOA1 and ABCA1 in the ileum of 8-week-old mice ( n = 5). (J) Relative mRNA levels of Apoa1 , Pon1 , and Pon3 in the ileum of 8-week-old mice ( n = 5). (K) Serum APOA1 levels in 8-, 16- and 32-week-old mice ( n = 8–10). (L) Serum APOA1 levels in 8-week-old mice exposed to HFCS for 14 days ( n = 8). (M) Serum APOA1 levels in 16-week-old Pparα Δhep mice ( n = 10). (N) Schematic representation of a cocktail of broad-spectrum antibiotics (Abx) experimental design. (O) Representative images stained with H&E and Oil Red O, and immunofluorescent staining for F4/80 (red) in the liver from 8-week-old mice treated with Abx ( n = 5). (P) Triglyceride in serum and liver treated with Abx ( n = 10). (Q) Relative mRNA levels of genes related to triglyceride accumulation in the liver from 8-week-old mice treated with Abx ( n = 5). (R) Hepatic levels of cytokines from 8-week-old mice treated with Abx ( n = 5). (S) Protein levels of F4/80 in the liver of 8-week-old mice treated with Abx ( n = 3). (T) Relative mRNA levels of F4/80, Clec4f , and Cd14 in the liver of 8-week-old mice treated with Abx ( n = 5). LD: lipid droplet; M: mitochondria; FITC-LPS: fluorescein isothiocyanate (FITC)-LPS; Au-LPS: LPS-gold-complexes. Data are shown as the mean ± SD. An unpaired two-tailed Student's t -test; ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: These samples were incubated overnight with a polyclonal rabbit anti-mouse F4/80 (1:1000, #A1256, ABclonal), polyclonal rabbit anti-mouse APOA1 (1:2000, #A14211, ABclonal), polyclonal rabbit anti-mouse ABCA1 (1:1000, #NB400-105, Novus Biological, Centennial), monoclonal rabbit anti-mouse CD14 (1:1000, #A19011, ABclonal), or polyclonal rabbit anti-mouse PV1 (1:2000, #A15906, ABclonal).

Techniques: Translocation Assay, Derivative Assay, Permeability, Sequencing, Transmission Assay, Electron Microscopy, Fluorescence, Microscopy, Staining, Two Tailed Test

Journal: The Journal of Clinical Investigation

Article Title: LXR signaling pathways link cholesterol metabolism with risk for prediabetes and diabetes

doi: 10.1172/JCI173278

Figure Lengend Snippet: MESA participants with lower expression of LXRα target genes, ABCA1 , ABCG1 , and MYLIP (lowest tertile), in 2 independent sub-studies, the Initial Study ( A ) and the Replication Study ( B ), were more likely to develop prediabetes/T2D compared with those with greater expression (highest tertile). Cox proportional hazards regression models were used, adjusting for age, sex, race and ethnicity, cigarette smoking, physical activity level, BMI, triglycerides, HDL-cholesterol, and systolic blood pressure (SBP) in the full model. The x axis is in logarithmic scale.

Article Snippet: After blocking in 5% blocking buffer (Bio-Rad, catalog 1706404) for 1 hour at room temperature, the membranes were incubated with primary antibodies in 3% BSA overnight at 4°C: rabbit anti-ABCG1 (Abcam, catalog ab52617; diluted 1:1,000), rabbit anti-MYLIP (Invitrogen, catalog PA5-96524; diluted 1:500), mouse anti-ABCA1 (Abcam, catalog ab1818; diluted 1:200), and mouse anti–β-actin (Sigma-Aldrich, catalog A5441; diluted 1:10,000).

Techniques: Expressing, Activity Assay

Journal: Stroke and Vascular Neurology

Article Title: Enhanced liver X receptor signalling reduces brain injury and promotes tissue regeneration following experimental intracerebral haemorrhage: roles of microglia/macrophages

doi: 10.1136/svn-2023-002331

Figure Lengend Snippet: GW3965 upregulates LXR downstream-gene expression. (A) Representative images of confocal tile scan showing whole day 7 lesion labelled with DAPI (blue), GFAP (Cyan), ABCA1 (green) and Iba1 (red). Scale bar, 200 µm. (B) Representative image of a three-dimensional (3D) reconstruction of a perihaematomal region in the treatment group using Imaris. Scale bar, 3 µm. (C) Representative confocal images of perihaematomal region labelled with DAPI (blue), GFAP (cyan), ABCA1 (green) and Iba1 (red). The left corner within dotted lines depicts lesion centre. Scale bar, 100 µm. (D–F), Bar graphs comparing the ABCA1 + percentage (D), Iba1 + ABCA1 + over Iba1 + proportions (E) and GFAP + ABCA1 + over GFAP + proportions (F) between GW3965 treatment and DMSO control within the lesion region at day 7; the sample size was n=10 per group. (G) Representative images of confocal tile scan showing whole day 7 lesion labelled with DAPI (blue), Iba1 (cyan), GFAP (green) and ApoE (red). Scale bar, 200 µm. Scale bars are not equal. (H), Representative image of a 3D reconstruction of a perihaematomal region in the treatment group using Imaris. Scale bar, 7 µm. (I), Representative confocal images of perihaematomal region labelled with DAPI (blue), Iba1 (cyan), GFAP (green) and ApoE (red). The left corner within dotted lines is the lesion centre. Scale bar, 100 µm. (J–L), Bar graphs comparing the ApoE + percentage (J), GFAP + ApoE + over GFAP + proportions (K) and Iba1 + ApoE + over Iba1 + proportions (L) between GW3965 treatment and DMSO control within the lesion region at day 7; the sample size was n=10 per group. Significance is indicated as *p<0.05, **p<0.01, ****p<0.0001; two-tailed, unpaired t-test (D, F, K); or two-tailed, unpaired t-test with Welch’s correction (E); or Mann-Whitney U test (J, L). Bar graphs show individual data points and represent mean±SD. ApoE, apolipoprotein E; LXR, liver X receptor.

Article Snippet: After transferring to Hybond polyvinylidene difluoride (Amersham) membrane using a wet transfer system (Bio-Rad), membranes were blocked with 5% BSA and then probed overnight with the following primary antibodies: rabbit anti-human/mouse ABCA1 (1:1000; Cell Signaling, 96292), rabbit anti-mouse ApoE (1:1000; Cell Signaling, 49285), mouse anti-human/mouse IL-1β (1:1000; Cell Signaling, 12242), rabbit anti-mouse iNOS (1:1000; Cell Signaling, 13120), rabbit anti-human/mouse CD206 (1:1000; Cell Signaling, 24595), rabbit anti-human/mouse Arginase 1 (1:1000; Cell Signaling, 93668) and rabbit anti-mouse β-Actin (1:1000; Cell Signaling, 24595).

Techniques: Gene Expression, Control, Two Tailed Test, MANN-WHITNEY