Journal: PLoS ONE

Article Title: Oxidation by Neutrophils-Derived HOCl Increases Immunogenicity of Proteins by Converting Them into Ligands of Several Endocytic Receptors Involved in Antigen Uptake by Dendritic Cells and Macrophages

doi: 10.1371/journal.pone.0123293

Figure Lengend Snippet: ( A ) Uptake of fluorescently-labelled proteins by CHO cells transfected with human CD36 as compared to non-transfected cells. ( B ) Binding of anti-CD36 mAb and control mouse IgA to CD36-transfected and non-transfected CHO cells, determined by cellular ELISA. ( C ) Binding of rCD36 to plate-adsorbed proteins. ( D ) Effects of 10 or 100 μg/ml of indicated, soluble ligands on rCD36 binding to plate-adsorbed OVA-Cl. ( E ) Binding of polyclonal anti-mouse CD36 Ab and control goat IgG to BM-DC, splenic DC and PEM. A representative histogram of Ab binding to BM-DC is displayed on the left graph. ( F ) Binding of anti-mouse SR-A mAb and control rat IgG2b to BM-DC, splenic DC and PEM. A representative histogram of Ab binding to BM-DC is shown on the left graph. ( G ) Binding of SR-A present in lysates of PEM to plate-adsorbed proteins. ( H ) Effects of anti-SR-A 2F8 mAb and AcLDL on the uptake of fluorescently-labelled proteins by BM-DC. Shown are results of single experiments, each representative of at least 3 similar experiments performed (A-D, G, H) or averages +SEM from 4–6 independent experiments (E, F). The data were analysed with the unpaired (A-C, G) or one-sample (H) Student’s t-test or with ANOVA, followed by the Tukey-Kramer post-test (D). *, p < 0.05; ND, not done.

Article Snippet: Rat anti-mouse scavenger receptor A (SR-A) mAb (clone 2F8) was obtained from AbD Serotec; mouse anti-mouse CD36 mAb (CRF D-2712) from Hycult Biotech; mouse IgA isotype control mAb (M18-254), rat IgG2b isotype control mAb (A95-1), rat anti-mouse CD11b mAb (M1/70) and phycoerythrin (PE)-streptavidin conjugate from BD Biosciences; rat IgG2a isotype control mAb (54447), normal goat IgG, polyclonal goat anti-mouse CD36, anti-mouse LOX-1 (lectin-type oxidised LDL receptor-1), anti-human SREC-I (scavenger receptor expressed by endothelial cells-I) Ab and PE-conjugated rat anti-mouse LOX-1 mAb (214012) from R&D Systems; polyclonal goat anti-mouse SREC-I, anti-mouse RAGE (receptor for advanced glycation end products), anti-mouse stabilin-1 and rabbit anti-mouse-stabilin-1 Ab from Santa Cruz Biotechnology; PE-conjugated donkey anti-goat IgG Ab from SouthernBiotech; rat anti-mouse CD206/mannose receptor mAb (MR5D3) and PE-conjugated goat anti-rat IgG Ab from BioLegend; horseradish peroxidase (HRP)-conjugated rabbit anti-mouse IgA, F(ab’)2 fragments of goat anti-rat IgG and donkey anti-goat IgG Ab from Rockland.

Techniques: Transfection, Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: Cell Communication and Signaling : CCS

Article Title: SIRT3 deficiency exacerbates fatty liver by attenuating the HIF1α-LIPIN 1 pathway and increasing CD36 through Nrf2

doi: 10.1186/s12964-020-00640-8

Figure Lengend Snippet: Enhanced hepatic VLDLR and CD36 levels in Sirt3 -deficient mice fed a HFD. mRNA abundance ( a ) and protein levels ( b ) of VLDLR in livers of WT and Sirt3 −/− mice fed either a standard chow or a HFD. CD36 ( c ), total and phospho-Nrf2 ( d ), Keap1 ( e ) and NQO1 ( f ) protein levels. g , ROS levels. h , PPARγ protein levels. Data are presented as the mean ± S.D. ( n = 6 per group). a, p < 0.05 vs. WT mice fed a standard chow. b, p < 0.05 vs. WT mice fed a HFD. c, p < 0.05 vs. Sirt3 −/− mice fed a standard chow

Article Snippet: Western blot analysis was performed using antibodies against HIF1α (sc-10,790), LIPIN1 (sc-98,450), Histone H3 (sc-10,809), SREBP1 (sc-365,513), Nrf2 (sc-722), NQO1 (sc-393,736), PPARγ (sc-7273) (Santa Cruz Biotechnology), SIRT3 (#5490), phospho-mTOR Ser2481 (#2974), mTOR (#2972), Keap1 (#4678 s) (Cell Signaling Technology Inc., Danvers, MA), GAPDH (MAB374) (Merck Millipore), CD36 (NB400–144) (Novus Biologicals, Centennial, CO), VLDLR (AF2258) (R&D Systems, Minneapolis, MN).

Techniques:

Journal: Cell Communication and Signaling : CCS

Article Title: SIRT3 deficiency exacerbates fatty liver by attenuating the HIF1α-LIPIN 1 pathway and increasing CD36 through Nrf2

doi: 10.1186/s12964-020-00640-8

Figure Lengend Snippet: The increase in CD36 levels caused by lipids in Sirt3 -deficient hepatocytes is mediated by Nrf2. VLDLR mRNA abundance ( a ) and protein levels of VLDLR and NQO1, an Nrf-2-target gene, ( b ) were assessed in Huh-7 cells incubated with fatty acid free-BSA or BSA-palmitate (0.3 mM) and exposed to either vehicle or the Sirt3 inhibitor AAPBO (100 μM) for 16 h. a, p < 0.05 vs. CT. b, p < 0.05 vs. CT cells incubated with palmitate. c, p < 0.05 vs. CT cells treated with AAPBO. c , fatty acid uptake in Huh-7 cells incubated with fatty acid free-BSA or BSA-palmitate (0.3 mM) and exposed to either vehicle or the Sirt3 inhibitor AAPBO (100 μM) for 16 h was measured by the uptake of BODIPY-C16. a, p < 0.05 vs. CT. b, p < 0.05 vs. CT cells incubated with palmitate. c, p < 0.05 vs. CT cells treated with AAPBO. mRNA abundance ( d ) and protein levels of VLDLR ( e ) in Huh-7 cells transfected with control (CT) or SIRT3 siRNA and incubated in the presence or absence 0.3 mM palmitate (Pal) for 24 h. Protein levels of CD36 ( f ), NQO1 ( g ) and PPARγ ( h ) in Huh-7 cells transfected with control (CT) or SIRT3 siRNA and incubated in the presence or absence 0.3 mM palmitate (Pal) or the Nrf2 inhibitor ML385 (10 μM) for 24 h. a, p < 0.05 vs. CT siRNA cells. b, p < 0.05 vs. CT siRNA cells incubated with palmitate. c, p < 0.05 vs. SIRT3 siRNA cells. d, p < 0.05 vs. CT siRNA cells incubated with palmitate and ML385

Article Snippet: Western blot analysis was performed using antibodies against HIF1α (sc-10,790), LIPIN1 (sc-98,450), Histone H3 (sc-10,809), SREBP1 (sc-365,513), Nrf2 (sc-722), NQO1 (sc-393,736), PPARγ (sc-7273) (Santa Cruz Biotechnology), SIRT3 (#5490), phospho-mTOR Ser2481 (#2974), mTOR (#2972), Keap1 (#4678 s) (Cell Signaling Technology Inc., Danvers, MA), GAPDH (MAB374) (Merck Millipore), CD36 (NB400–144) (Novus Biologicals, Centennial, CO), VLDLR (AF2258) (R&D Systems, Minneapolis, MN).

Techniques: Incubation, Transfection, Control

Journal: Cell Communication and Signaling : CCS

Article Title: SIRT3 deficiency exacerbates fatty liver by attenuating the HIF1α-LIPIN 1 pathway and increasing CD36 through Nrf2

doi: 10.1186/s12964-020-00640-8

Figure Lengend Snippet: Potential new mechanisms by which Sirt3 deficiency promotes hepatic steatosis in mice fed a HFD. Exposure to a HFD reduces Sirt3 and contributes to triglyceride accumulation in the liver. However, hepatic lipid accumulation is attenuated by the activation of an adaptive mechanism involving an increase in nuclear HIF-1α and Lipin 1. Longer exposures to a HFD exacerbates Sirt3 decrease leading to higher uptake of lipids through an Nrf-2 mediated increase in CD36 and VLDLR. This results in a higher increase in fatty acid accumulation, which in turn reduces succinate levels, ultimately suppressing the adaptive increase in HIF-1α and Lipin 1. FAO: Fatty acid oxidation

Article Snippet: Western blot analysis was performed using antibodies against HIF1α (sc-10,790), LIPIN1 (sc-98,450), Histone H3 (sc-10,809), SREBP1 (sc-365,513), Nrf2 (sc-722), NQO1 (sc-393,736), PPARγ (sc-7273) (Santa Cruz Biotechnology), SIRT3 (#5490), phospho-mTOR Ser2481 (#2974), mTOR (#2972), Keap1 (#4678 s) (Cell Signaling Technology Inc., Danvers, MA), GAPDH (MAB374) (Merck Millipore), CD36 (NB400–144) (Novus Biologicals, Centennial, CO), VLDLR (AF2258) (R&D Systems, Minneapolis, MN).

Techniques: Activation Assay

Journal: Cardiovascular research

Article Title: Beneficial effects of combinatorial micronutrition on body fat and atherosclerosis in mice.

doi: 10.1093/cvr/cvr146

Figure Lengend Snippet: Figure 2 Lipistase enhances muscle lipid utilization in LDLrKO mice. (A) Representative images of anti-CD36 immunostaining on paraffin-embedded sections of oxidative muscle (gastrocnemius) from mice after 10-month lipistase treatment vs. controls (bars ¼ 50 mm). (B) Oxygen consumption measured after the 10-month treatment (***P , 0.001). (C) Immunoblot quantification of the membrane-bound CD36 protein in muscle and mRNA expression of marker genes for fatty acid transport and oxidation measured in gastrocnemius 10 months after lipistase treatment (*P , 0.05).

Article Snippet: For CD36 quantification by immunoblotting, 4 mg and 6 mg of cytosolic and membrane extracts, respectively, were loaded and probed with 1/1000 rabbit polyclonal anti-CD36 antibody (Novus Biologicals, Inc. Littleton, CO, USA).

Techniques: Immunostaining, Western Blot, Membrane, Expressing, Marker

Journal: Circulation: Cardiovascular Genetics

Article Title: CD36 Is a Matrix Metalloproteinase-9 Substrate That Stimulates Neutrophil Apoptosis and Removal During Cardiac Remodeling

doi: 10.1161/circgenetics.115.001249

Figure Lengend Snippet: Figure 3: MMP-9 regulates CD36 levels through proteolytic degradation. (A) Full length CD36

Article Snippet: After blocking with 5% nonfat milk (Bio-Rad), the membrane was probed with primary antibodies: rabbit anti-CD36 polyclonal antibody (Novus BiologicalsNB400-144) at 1:1000, goat anti-CILP antibody (Aviva Systems Biology, OAEB01146) at 1:1000, or rabbit anti-caspase-3 (Cell Signaling, 9662) at

Techniques:

Journal: Circulation: Cardiovascular Genetics

Article Title: CD36 Is a Matrix Metalloproteinase-9 Substrate That Stimulates Neutrophil Apoptosis and Removal During Cardiac Remodeling

doi: 10.1161/circgenetics.115.001249

Figure Lengend Snippet: Figure 2: MMP-9 deletion increased CD36 and CILP levels at day 7 post-myocardial infarction

Article Snippet: After blocking with 5% nonfat milk (Bio-Rad), the membrane was probed with primary antibodies: rabbit anti-CD36 polyclonal antibody (Novus BiologicalsNB400-144) at 1:1000, goat anti-CILP antibody (Aviva Systems Biology, OAEB01146) at 1:1000, or rabbit anti-caspase-3 (Cell Signaling, 9662) at

Techniques:

Journal: Circulation: Cardiovascular Genetics

Article Title: CD36 Is a Matrix Metalloproteinase-9 Substrate That Stimulates Neutrophil Apoptosis and Removal During Cardiac Remodeling

doi: 10.1161/circgenetics.115.001249

Figure Lengend Snippet: Figure 5: MMP-9 regulates phagocytosis through CD36. As shown by the in vivo stimulated

Article Snippet: After blocking with 5% nonfat milk (Bio-Rad), the membrane was probed with primary antibodies: rabbit anti-CD36 polyclonal antibody (Novus BiologicalsNB400-144) at 1:1000, goat anti-CILP antibody (Aviva Systems Biology, OAEB01146) at 1:1000, or rabbit anti-caspase-3 (Cell Signaling, 9662) at

Techniques: In Vivo

Journal: Circulation: Cardiovascular Genetics

Article Title: CD36 Is a Matrix Metalloproteinase-9 Substrate That Stimulates Neutrophil Apoptosis and Removal During Cardiac Remodeling

doi: 10.1161/circgenetics.115.001249

Figure Lengend Snippet: Figure 8: Diagram depicting the roles of MMP-9 mediated CD36 degradation in LV remodeling

Article Snippet: After blocking with 5% nonfat milk (Bio-Rad), the membrane was probed with primary antibodies: rabbit anti-CD36 polyclonal antibody (Novus BiologicalsNB400-144) at 1:1000, goat anti-CILP antibody (Aviva Systems Biology, OAEB01146) at 1:1000, or rabbit anti-caspase-3 (Cell Signaling, 9662) at

Techniques:

Journal: Frontiers in Endocrinology

Article Title: The Effect of Hypoxia and Metformin on Fatty Acid Uptake, Storage, and Oxidation in L6 Differentiated Myotubes

doi: 10.3389/fendo.2018.00616

Figure Lengend Snippet: Protein expression of FFA protein transporters. The effect of mild (4% O 2 ) and severe (1% O 2 ) hypoxia (A) and metformin administration (B) on CD36 and SCL27A4 protein expression. * p < 0.05 for comparison with control exposures (20% O 2 , unpaired T -test), # p < 0.05 for comparison with vehicle-treated group (unpaired T -test), n = 6–9.

Article Snippet: Western blotting: Protein expression of SLC27A4, CD36, and GAPDH (Glyceraldehyde 3-phosphate dehydrogenase—a loading control) was performed by western blot analysis using rabbit polyclonal antibodies (Anti-SLC27A4 antibody: ab199718, Anti-CD36 antibody: ab133625, and Anti-GAPDH antibody: ab9485, all from Abcam, Cambridge, UK).

Techniques: Expressing

Journal: Frontiers in Endocrinology

Article Title: The Effect of Hypoxia and Metformin on Fatty Acid Uptake, Storage, and Oxidation in L6 Differentiated Myotubes

doi: 10.3389/fendo.2018.00616

Figure Lengend Snippet: Gene expression of FFA protein transporters and key regulators of mitochondria metabolism. The effect of mild (4% O 2 ) and severe (1% O 2 ) hypoxia (A) and metformin administration (B) on CD36, SCL27A4, CPT1, UCP2, and PGC1α gene expression. * p < 0.05 for comparison with control exposures (20% O 2 ), # p < 0.05 for comparison with vehicle-treated group, n = 6. Gene expression data were analyzed using the REST Software (Qiagen, Hilden, Germany) based on the Pfaffl's method.

Article Snippet: Western blotting: Protein expression of SLC27A4, CD36, and GAPDH (Glyceraldehyde 3-phosphate dehydrogenase—a loading control) was performed by western blot analysis using rabbit polyclonal antibodies (Anti-SLC27A4 antibody: ab199718, Anti-CD36 antibody: ab133625, and Anti-GAPDH antibody: ab9485, all from Abcam, Cambridge, UK).

Techniques: Expressing, Software

Journal: Journal of Inflammation Research

Article Title: Catalpol Alleviates HFpEF via Inhibition of the S100A8-RAGE-NOX4 Inflammatory Axis in Murine Hearts

doi: 10.2147/JIR.S552741

Figure Lengend Snippet: Visualization of molecular docking results using interaction diagrams. ( A ) Molecular docking of CAT and ERK1. ( B ) Molecular docking of CAT and MMP2. ( C ) Molecular docking of CAT and NOX4. ( D ) Molecular docking of CAT and RAGE. ( E ) Molecular docking of CAT and S100A8.

Article Snippet: Primary antibodies against the following targets were used: S100A8 (15792-1-AP), RAGE (16346-1-AP), NOX4 (14347-1-AP), MMP2 (10373-2-AP), and GAPDH (60004-1-AP) from Proteintech (Wuhan, China); ERK1/2 (9102), phospho-ERK1/2 (p-ERK1/2, 4370), p65 (8242), and phospho-p65 (p-p65, 3033) from Cell Signaling Technology (Danvers, MA, USA).

Techniques:

Journal: Journal of Inflammation Research

Article Title: Catalpol Alleviates HFpEF via Inhibition of the S100A8-RAGE-NOX4 Inflammatory Axis in Murine Hearts

doi: 10.2147/JIR.S552741

Figure Lengend Snippet: CAT downregulated the S100A8-RAGE-NOX4 signaling pathway in HFpEF. ( A ) Representative WB bands of S100A8, RAGE, NOX4, ERK, p-ERK, P65, p-P65, and MMP2. ( B – G ) Quantification of protein expression levels analyzed using ImageJ software (n=3). ( H ) Quantitative analysis of IL-1β expression (n=3). ( I ) Quantitative analysis of IL-6 expression (n=3). ( J ) Quantitative analysis of TNF-α expression (n=3). Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Primary antibodies against the following targets were used: S100A8 (15792-1-AP), RAGE (16346-1-AP), NOX4 (14347-1-AP), MMP2 (10373-2-AP), and GAPDH (60004-1-AP) from Proteintech (Wuhan, China); ERK1/2 (9102), phospho-ERK1/2 (p-ERK1/2, 4370), p65 (8242), and phospho-p65 (p-p65, 3033) from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Software