Journal: bioRxiv

Article Title: Macrophage scavenger receptor 1 controls Chikungunya virus infection through autophagy

doi: 10.1101/2020.02.28.969832

Figure Lengend Snippet: 8 weeks old, sex-matched, C57BL/6 (WT) and Msr1 deficient ( Msr1 -/- ) mice were infected with CHIKV. Quantitative RT-PCR analyses of A ) Msr1 , CHIKV, type I IFN, ISG mRNA (N=4), B ) CHIKV loads in whole blood cells of WT and Msr1 -/- mice (N=8 per genotype). C ) Viremia presented as plaque forming units (PFU) per ml serum (N=8 per genotype). D ) Quantitative RT-PCR analyses of CHIKV loads in the ankle joints (N=8 per genotype). E ) Fold changes in the footpad dimensions of infected mice over uninfected (day 0) (N=8 per experimental group). F ) Representative micrographs of hematoxylin and eosin staining of ankle joints at 8 days after infection. N=5/genotype. B: bone, T: tendon, M: muscle. Magnification: 200x. G ) Arbitrary scores of ankle joint inflammation and damage using scales of 1 to 5, with 5 representing the worst disease presentation. Each dot=one mouse, the small horizontal line: the median of the result. N=5 per genotype. The data represent two independent experiments. *, P<0.05; **, p<0.01; ***, P<0.001 [ B-D ) non-parametric t-test, E, G ) Student’s t-test].

Article Snippet: The Myc-DDK-tagged mouse Msr1 ORF clone (Cat# MR205384), Atg12 (Cat# MR200886), Atg5 (Cat# MR203691), Atg16l1 (Cat# MR209513), Atg9a (Cat# MR208753), Atg13 (Cat# MR207683) and, p62/Sqstm1 (Cat# MR226105), human MSR1 (Cat# RC209609) and Beclin 1 (Cat# MR207162) were obtained from Origene (Rockville, MD 20850, USA). pcDNA-FLAG-Ulk1 (Plasmid # 27636) and pcDNA4-VPS34-FLAG (Plasmid # 24398) were purchased from Addgene (Cambridge, MA 02139, USA).

Techniques: Infection, Quantitative RT-PCR, Staining

Journal: bioRxiv

Article Title: Macrophage scavenger receptor 1 controls Chikungunya virus infection through autophagy

doi: 10.1101/2020.02.28.969832

Figure Lengend Snippet: A ) Quantitative RT-PCR analyses of the cellular viral loads in bone marrow-derived macrophages (BMDM) infected with CHIKV at a multiplicity of infection (MOI) of 10. Bars: mean + s.e.m, n=4, * P<0.05 (Student’s t-test). B ) Microscopic images of immunofluorescence staining for LC3B in BMDMs infected with CHIKV at a MOI of 10 for 12hrs. C ) The mean number of LC3B-positive punctae per cell (n=30 cells per field x 3 fields). D ) Immunoblots of LC3B in BMDMs infected with CHIKV at a MOI of 10, following-treatment without/with 20µM of chloroquine (CQ) for 3hrs. E ) Generation of human ATG12 -/- and MSR1 -/- trophoblasts by CRISPR-Cas9. Both anti-ATG12 and ATG5 antibodies detect the stable ATG5-ATG12 dimer, which is abolished in ATG12 -/- cells. Monomeric ATG5 is liberated and detected in ATG12 -/- cells by an anti-ATG5 antibody. F ) Quantitative RT-PCR analyses of intracellular CHIKV RNA in trophoblasts infected with CHIKV at a MOI of 1. Bars: mean + s.e.m, n=2, **, P<0.01 (two-way ANOVA). G ) Immunoblots of LC3B in trophoblasts infected with CHIKV at a MOI of 1.

Article Snippet: The Myc-DDK-tagged mouse Msr1 ORF clone (Cat# MR205384), Atg12 (Cat# MR200886), Atg5 (Cat# MR203691), Atg16l1 (Cat# MR209513), Atg9a (Cat# MR208753), Atg13 (Cat# MR207683) and, p62/Sqstm1 (Cat# MR226105), human MSR1 (Cat# RC209609) and Beclin 1 (Cat# MR207162) were obtained from Origene (Rockville, MD 20850, USA). pcDNA-FLAG-Ulk1 (Plasmid # 27636) and pcDNA4-VPS34-FLAG (Plasmid # 24398) were purchased from Addgene (Cambridge, MA 02139, USA).

Techniques: Quantitative RT-PCR, Derivative Assay, Infection, Immunofluorescence, Staining, Western Blot, CRISPR

Journal: bioRxiv

Article Title: Macrophage scavenger receptor 1 controls Chikungunya virus infection through autophagy

doi: 10.1101/2020.02.28.969832

Figure Lengend Snippet: A ) Co- immunoprecipitation (co-IP) of FLAG-tagged mouse autophagy proteins with Myc-Msr1 expressed in HEK293 cells using a mouse monoclonal anti-FLAG antibody, followed by immunoblotting (IB) with a mouse monoclonal anti-FLAG and rabbit anti-Msr1 antibody. B ) co-IP of FLAG-Msr1 (mouse) with endogenous human ATG5-ATG12 complex expressed in HEK293. CHIKV infection: MOI of 1. WCE: whole cell extract. C ) co-IP of endogenous Msr1 with the Atg5-Atg12 complex from bone marrow-derived macrophages (BMDMs) following CHIKV infection at a MOI of 10 for 12 and 24hrs. The IP was carried out with a rabbit anti-Msr1 antibody cross-linked to protein A/G agarose beads. Msr1 -/- cell serves as a negative control. D ) Microscopic images of immunofluorescence staining for Atg12 and Msr1 in BMDMs infected without (mock) /with CHIKV at a MOI of 10 for 12hrs. Atg12 and Msr1 were stained by a mouse anti-Atg12 and rabbit anti-Msr1, followed by an Alexa Fluor-488 (green) and -594 (red)-conjugated secondary antibody respectively. The cells were counterstained for nuclei by DAPI (blue). The yellow punctae in the overlay indicate co-localizations. Magnification: 400x. E ) co-IP of FLAG-MSR1 (human) or its fragment (aa1-50, 51-end) with endogenous ATG5-ATG12 complex expressed in HEK293 cells. CHIKV infection: MOI of 1 for 12hrs. F ) Microscopic images of immunofluorescence staining for LC3B and MSR1 in trophoblasts without (mock) / infected with CHIKV at a MOI of 0.5 for 12hrs. The yellow punctae in the overlay indicate co-localizations. Magnification: 400x.

Article Snippet: The Myc-DDK-tagged mouse Msr1 ORF clone (Cat# MR205384), Atg12 (Cat# MR200886), Atg5 (Cat# MR203691), Atg16l1 (Cat# MR209513), Atg9a (Cat# MR208753), Atg13 (Cat# MR207683) and, p62/Sqstm1 (Cat# MR226105), human MSR1 (Cat# RC209609) and Beclin 1 (Cat# MR207162) were obtained from Origene (Rockville, MD 20850, USA). pcDNA-FLAG-Ulk1 (Plasmid # 27636) and pcDNA4-VPS34-FLAG (Plasmid # 24398) were purchased from Addgene (Cambridge, MA 02139, USA).

Techniques: Immunoprecipitation, Co-Immunoprecipitation Assay, Western Blot, Infection, Derivative Assay, Negative Control, Immunofluorescence, Staining

Journal: bioRxiv

Article Title: Macrophage scavenger receptor 1 controls Chikungunya virus infection through autophagy

doi: 10.1101/2020.02.28.969832

Figure Lengend Snippet: A, B ) Repression of CHIKV replication by MSR1 requires ATG5. Mouse primary embryonic fibroblasts were transfected with either 0.2 or 1.5 µg of empty vector (Vec) or FLAG-Msr1 (mouse) expression plasmid. 24hrs later the cells were then infected with CHIKV (multiplicity of infection MOI=0.5). A ) Immunoblots showing FLAG-Msr1 (mouse) and Atg5-Atg12 expression at 24hrs after plasmid DNA transfection. B ) Quantitative RT-PCR analyses of intracellular CHIKV RNA. C ) Repression of CHIKV replication by MSR1 requires ATG12. Human trophoblasts were transfected with either 0.2µg of empty vector or a FLAG-MSR1 (human) expression plasmid. Twenty four hours later, the cells were then infected with CHIKV MOI=0.5 for 12hrs. Intracellular CHIKV RNA was quantitated by RT-PCR. D ) Generation of human ATG16L1 -/- trophoblasts by CRISPR-Cas9. The immunoblot shows ATG16L1 knockout efficiency. E ) Repression of CHIKV replication by MSR1 requires the FBD domain of ATG16L1. ATG16L1 -/- trophoblasts were transfected with the indicated combinations of expression plasmids (human gene) and vector (Vec) (50ng each) respectively. After 24hrs, the cells were infected with CHIKV at a MOI of 0.5 for 16hrs. FL: full-length, ΔFBD: FBD domain deletion, ΔWD40: WD40 domain deletion of ATG16L1. The immunoblots show CHIKV Capsid and cellular protein expression. Bars: mean + s.e.m, *, P<0.05, **, P<0.01 (Student’s t-test), N=3.

Article Snippet: The Myc-DDK-tagged mouse Msr1 ORF clone (Cat# MR205384), Atg12 (Cat# MR200886), Atg5 (Cat# MR203691), Atg16l1 (Cat# MR209513), Atg9a (Cat# MR208753), Atg13 (Cat# MR207683) and, p62/Sqstm1 (Cat# MR226105), human MSR1 (Cat# RC209609) and Beclin 1 (Cat# MR207162) were obtained from Origene (Rockville, MD 20850, USA). pcDNA-FLAG-Ulk1 (Plasmid # 27636) and pcDNA4-VPS34-FLAG (Plasmid # 24398) were purchased from Addgene (Cambridge, MA 02139, USA).

Techniques: Transfection, Plasmid Preparation, Expressing, Infection, Western Blot, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, CRISPR, Knock-Out

Journal: bioRxiv

Article Title: Macrophage scavenger receptor 1 controls Chikungunya virus infection through autophagy

doi: 10.1101/2020.02.28.969832

Figure Lengend Snippet: Co-immunoprecipitation (co-IP) of FLAG-Msr1 (mouse) with Myc-tagged CHIKV proteins expressed in HEK293 cells using a mouse anti-FLAG antibody, followed by immunoblotting (IB) with a rabbit anti-FLAG and anti-Myc antibody. WCE: whole cell extract. N.S: non-specific bands. E: CHIKV envelope, C: capsid, P: non-structural protein. C ) co-IP of FLAG-MSR1 (human) with Myc-nsP1 in HEK293. D ) Microscopic images of immunofluorescence staining for endogenous MSR1 and CHIKV nsP1 in trophoblasts without (mock) / infected with CHIKV at a multiplicity of infection of 0.5 for 12hrs. MSR1 and nsP1 were stained by a rabbit anti-MSR1 and rat anti-nsP1, followed by an Alexa Fluor-594 (red) and -488 (green) conjugated secondary antibody respectively. The cells were counterstained for nuclei by DAPI (blue). Scale bar: 20µM. The yellow punctae in the overlay indicate co-localizations.

Article Snippet: The Myc-DDK-tagged mouse Msr1 ORF clone (Cat# MR205384), Atg12 (Cat# MR200886), Atg5 (Cat# MR203691), Atg16l1 (Cat# MR209513), Atg9a (Cat# MR208753), Atg13 (Cat# MR207683) and, p62/Sqstm1 (Cat# MR226105), human MSR1 (Cat# RC209609) and Beclin 1 (Cat# MR207162) were obtained from Origene (Rockville, MD 20850, USA). pcDNA-FLAG-Ulk1 (Plasmid # 27636) and pcDNA4-VPS34-FLAG (Plasmid # 24398) were purchased from Addgene (Cambridge, MA 02139, USA).

Techniques: Immunoprecipitation, Co-Immunoprecipitation Assay, Western Blot, Immunofluorescence, Staining, Infection

Journal: bioRxiv

Article Title: Macrophage scavenger receptor 1 controls Chikungunya virus infection through autophagy

doi: 10.1101/2020.02.28.969832

Figure Lengend Snippet: A ) Co-immunoprecipitation (co-IP) of FLAG-Atg12 with Myc-Msr1 (mouse) proteins in the presence of nsP1 in HEK293T cells using a mouse monoclonal anti-FLAG antibody, followed by immunoblotting (IB) with a rabbit anti-FLAG or Msr1 antibody. WCE: whole cell extract. B ) Immunoblots of Myc-tagged CHIKV proteins after rapamycin (Rapa) treatment. Myc-tagged CHIKV gene expression plasmids were transfected into HEK293T cells for 24hrs. The cells were then treated with 100nM rapamycin with/without 40μM of chloroquine (CQ) for 24hrs. C ) The immunoblots show P62 knockout by CRISPR-Cas9 in human trophoblasts. D ) Immunoblots of Myc-tagged CHIKV proteins in trophoblasts after CHIKV infection. Myc-Caps, nsP1 expression plasmids were transfected into trophoblasts for 24hrs. The cells were then infected with CHIKV at a MOI of 0.5 for 12hrs.

Article Snippet: The Myc-DDK-tagged mouse Msr1 ORF clone (Cat# MR205384), Atg12 (Cat# MR200886), Atg5 (Cat# MR203691), Atg16l1 (Cat# MR209513), Atg9a (Cat# MR208753), Atg13 (Cat# MR207683) and, p62/Sqstm1 (Cat# MR226105), human MSR1 (Cat# RC209609) and Beclin 1 (Cat# MR207162) were obtained from Origene (Rockville, MD 20850, USA). pcDNA-FLAG-Ulk1 (Plasmid # 27636) and pcDNA4-VPS34-FLAG (Plasmid # 24398) were purchased from Addgene (Cambridge, MA 02139, USA).

Techniques: Immunoprecipitation, Co-Immunoprecipitation Assay, Western Blot, Gene Expression, Transfection, Knock-Out, CRISPR, Infection, Expressing

Journal: Scientific Reports

Article Title: CD206 + tumor-associated macrophages promote proliferation and invasion in oral squamous cell carcinoma via EGF production

doi: 10.1038/s41598-019-51149-1

Figure Lengend Snippet: Association of tumor-associated macrophages (TAMs) with clinicopathologic characteristics in OSCC.

Article Snippet: After rinsing, the cells were incubated at normal temperature for 20 min in dark with PE anti-human CD163 antibodies (Clone GHI/61, IgG1, κ ; BioLegend), FITC anti-human CD204 antibodies (Clone REA460, IgG1; Miltenyi Biotec, Bergisch, Gladbach, Germany), and PerCP/Cy5.5 anti-human CD206 antibodies (Clone 15-2, IgG1, κ ; BioLegend).

Techniques: Significance Assay

Journal: Scientific Reports

Article Title: CD206 + tumor-associated macrophages promote proliferation and invasion in oral squamous cell carcinoma via EGF production

doi: 10.1038/s41598-019-51149-1

Figure Lengend Snippet: Distribution of tumor-associated macrophages (TAMs) and epithelial growth factor (EGF) in oral squamous cell carcinoma (OSCC) patients. ( A ) Representative images of paraffin sections in/around tumors stained with H&E (a–c) and CD163 (d–f), CD204 (g–i), CD206 (j–l), and EGF (m–o) antibodies (brown). Counterstaining with Mayer’s hematoxylin is shown in blue. Scale bars, 100 μm. ( B ) Triple immunofluorescence staining was performed with CD163 (green), CD204 (red), and CD206 (white); nuclei were stained with DAPI (blue). Scale bars, 50 μm. ( C ) Double immunofluorescence staining performed with EGF (red) and CD163, CD204 or CD206 (green); nuclei were stained with DAPI (blue). Merged TAM markers and EGF images (yellow). The higher magnifications are displayed at the lower right. Scale bars, 50 μm.

Article Snippet: After rinsing, the cells were incubated at normal temperature for 20 min in dark with PE anti-human CD163 antibodies (Clone GHI/61, IgG1, κ ; BioLegend), FITC anti-human CD204 antibodies (Clone REA460, IgG1; Miltenyi Biotec, Bergisch, Gladbach, Germany), and PerCP/Cy5.5 anti-human CD206 antibodies (Clone 15-2, IgG1, κ ; BioLegend).

Techniques: Staining, Immunofluorescence, Double Immunofluorescence Staining

Journal: Scientific Reports

Article Title: CD206 + tumor-associated macrophages promote proliferation and invasion in oral squamous cell carcinoma via EGF production

doi: 10.1038/s41598-019-51149-1

Figure Lengend Snippet: Survival curves according to the expression of TAM subsets in OSCC. Survival rates were calculated by the Kaplan-Meier method with high versus low expression of CD163 + , CD204 + , or CD206 + TAMs. The classifications are described in the Materials and Methods section. Statistically significant differences between groups were determined by log-rank test ( A ) and ROC curve ( B ).

Article Snippet: After rinsing, the cells were incubated at normal temperature for 20 min in dark with PE anti-human CD163 antibodies (Clone GHI/61, IgG1, κ ; BioLegend), FITC anti-human CD204 antibodies (Clone REA460, IgG1; Miltenyi Biotec, Bergisch, Gladbach, Germany), and PerCP/Cy5.5 anti-human CD206 antibodies (Clone 15-2, IgG1, κ ; BioLegend).

Techniques: Expressing

Journal: Scientific Reports

Article Title: CD206 + tumor-associated macrophages promote proliferation and invasion in oral squamous cell carcinoma via EGF production

doi: 10.1038/s41598-019-51149-1

Figure Lengend Snippet: Univariate analysis of progression-free survival and disease-specific survival in advanced stage.

Article Snippet: After rinsing, the cells were incubated at normal temperature for 20 min in dark with PE anti-human CD163 antibodies (Clone GHI/61, IgG1, κ ; BioLegend), FITC anti-human CD204 antibodies (Clone REA460, IgG1; Miltenyi Biotec, Bergisch, Gladbach, Germany), and PerCP/Cy5.5 anti-human CD206 antibodies (Clone 15-2, IgG1, κ ; BioLegend).

Techniques:

Journal: The Journal of Biological Chemistry

Article Title: Recognition of lipoproteins by scavenger receptor class A members

doi: 10.1016/j.jbc.2021.100948

Figure Lengend Snippet: SCARA1 recognizes the modified LDL and VLDL. A , schematic models of the SR-A family members. B , FACS data of the hSCARA1-transfected HEK293 cells incubated with AcLDL, OxLDL, or LDL. Mock represents nontransfected cells. C , confocal fluorescent images of the hSCARA1 (with GFP tag)-transfected HEK293 cells incubated with LDL, AcLDL, or OxLDL (the scale bar represents 25 μm). D , FACS data of the hSCARA1-transfected HEK293 cells incubated with AcVLDL, OxVLDL, or VLDL. E , confocal fluorescent images of the hSCARA1 (with GFP-tag)-transfected HEK293 cells incubated with VLDL or OxVLDL (the scale bar represents 25 μm). F , FACS data of the hSCARA1-transfected HEK293 cells incubated with AcHDL, OxHDL, or HDL. G , confocal fluorescent images of the hSCARA1 (with mCherry tag)-transfected cells incubated with HDL, AcHDL, or OxHDL (the scale bar represents 25 μm). AcHDL, acetylated HDL; AcVLDL, acetylated VLDL; HDL, high-density lipoprotein; LDL, low-density lipoprotein; OxHDL, oxidized HDL; OxVLDL, oxidized VLDL; VLDL, very-low-density lipoprotein.

Article Snippet: The cDNAs of human SCARA1 to SCARA5 (GenBank: NM_138715.2, NM_006770, NM_016240.2, NM_130386 and NM_286133) were purchased from Sino Biological Inc or synthesized by Sangon Biotech.

Techniques: Modification, Transfection, Incubation

Journal: The Journal of Biological Chemistry

Article Title: Recognition of lipoproteins by scavenger receptor class A members

doi: 10.1016/j.jbc.2021.100948

Figure Lengend Snippet: SCARA1 recognizes the modified LDL and VLDL through its C-terminal SRCR domain. A , FACS data of the full-length hSCARA1 or the hSCARA1ΔSRCR-transfected HEK293 cells incubated with LDL or AcLDL. B , FACS data of the full-length hSCARA1 or the hSCARA1ΔSRCR-transfected HEK293 cells incubated with LDL or OxLDL. C , ELISA data of the interactions between the CL-SRCR fragment (homotrimer) and the SRCR domain (monomer) of hSCARA1 with the unmodified LDL or the modified LDL in the presence of Ca 2+ (2 mM). D , ELISA data of the interactions between the CL-SRCR fragment of hSCARA1 with the unmodified VLDL and the modified VLDL in the presence of Ca 2+ (2 mM). GFP was applied as a negative control. ELISA data are representative of three repeated experiments and presented as the mean ± SD. AcLDL, acetylated LDL; CL, collagen-like; LDL, low-density lipoprotein; OxLDL, oxidized LDL; SRCR, scavenger receptor cysteine-rich; VLDL, very-low-density lipoprotein.

Article Snippet: The cDNAs of human SCARA1 to SCARA5 (GenBank: NM_138715.2, NM_006770, NM_016240.2, NM_130386 and NM_286133) were purchased from Sino Biological Inc or synthesized by Sangon Biotech.

Techniques: Modification, Transfection, Incubation, Enzyme-linked Immunosorbent Assay, Negative Control

Journal: The Journal of Biological Chemistry

Article Title: Recognition of lipoproteins by scavenger receptor class A members

doi: 10.1016/j.jbc.2021.100948

Figure Lengend Snippet: MARCO and SCARA5 interact with the modified LDL and VLDL via the SRCR domains. A , FACS data for the binding of AcLDL with the hSCARA1-, hMARCO-, or hSCARA5-transfected cells. Mock represents nontransfected cells. B , FACS data for the binding of OxLDL with the hSCARA1-, hMARCO-, or hSCARA5-transfected cells. C , FACS data for the binding of AcVLDL or OxVLDL with the hSCARA1-, hMARCO-, or the hSCARA5-transfected cells. D , FACS data of the hMARCO- or hSCARA5-transfected cells incubated with AcHDL or OxHDL. E , confocal fluorescent images of the hMARCO (with GFP tag) transfected cells incubated with the modified or unmodified LDL or VLDL (the scale bar represents 25 μm). F , confocal fluorescent images of the hSCARA5 (with GFP tag)-transfected cells incubated with the modified or unmodified LDL or VLDL (the scale bar represents 25 μm). AcHDL, acetylated HDL; AcLDL, acetylated LDL; AcVLDL, acetylated VLDL; LDL, low-density lipoprotein; OxHDL, oxidized HDL; OxLDL, oxidized LDL; OxVLDL, oxidized VLDL; SRCR, scavenger receptor cysteine-rich; VLDL, very-low-density lipoprotein.

Article Snippet: The cDNAs of human SCARA1 to SCARA5 (GenBank: NM_138715.2, NM_006770, NM_016240.2, NM_130386 and NM_286133) were purchased from Sino Biological Inc or synthesized by Sangon Biotech.

Techniques: Modification, Binding Assay, Transfection, Incubation

Journal: The Journal of Biological Chemistry

Article Title: Recognition of lipoproteins by scavenger receptor class A members

doi: 10.1016/j.jbc.2021.100948

Figure Lengend Snippet: The interactions between the SRCR domains of SCARA1, MARCO, and SCARA5 with the modified LDL and VLDL are Ca 2+ dependent. A , ELISA data for the interactions of the SRCR domains of hSCARA1, hMARCO, or hSCARA5 with AcLDL in the presence of Ca 2+ (2 mM) or EDTA (1 mM). B , ELISA data for the interactions of the SRCR domains of hSCARA1, hMARCO, or hSCARA5 with OxLDL in the presence of Ca 2+ (2 mM) or EDTA (1 mM). C , ELISA data for the interactions of the SRCR domains of hSCARA1, hMARCO, or hSCARA5 with OxVLDL in the presence of Ca 2+ (2 mM) or EDTA (1 mM). D , FACS data of the hSCARA1-transfected cells incubated with the modified LDL or VLDL in the presence of Ca 2+ or EDTA. E , confocal fluorescent images of the hSCARA1 (with GFP-tag)-transfected cells incubated with the modified LDL or VLDL in the presence of EDTA (the scale bar represents 25 μm). F , FACS data of the hMARCO-transfected cells incubated with the modified LDL or VLDL in the presence of Ca 2+ or EDTA. G , FACS data of the hSCARA5-transfected cells incubated with the modified LDL or VLDL in the presence of Ca 2+ or EDTA. ELISA data are representative of three repeated experiments and presented as the mean ± SD. AcLDL, acetylated LDL; LDL, low-density lipoprotein; OxLDL, oxidized LDL; OxVLDL, oxidized VLDL; SRCR, scavenger receptor cysteine-rich; VLDL, very-low-density lipoprotein.

Article Snippet: The cDNAs of human SCARA1 to SCARA5 (GenBank: NM_138715.2, NM_006770, NM_016240.2, NM_130386 and NM_286133) were purchased from Sino Biological Inc or synthesized by Sangon Biotech.

Techniques: Modification, Enzyme-linked Immunosorbent Assay, Transfection, Incubation

Journal: The Journal of Biological Chemistry

Article Title: Recognition of lipoproteins by scavenger receptor class A members

doi: 10.1016/j.jbc.2021.100948

Figure Lengend Snippet: Comparison of the crystal structures of the SRCR domains of SCARA1, SCARA5, and MARCO. Crystal structures of the SRCR domains of hSCARA1 ( orange ), hSCARA5 ( blue , PDB ID:7C00), and mMARCO ( magenta , PDB ID: 2OY3) are shown as ribbon diagrams. The cation-binding sites ( dashed ovals ) of hSCARA1 and hSCARA5 are zoomed in at insets (Ca 2+ are shown as green spheres ; Mg 2+ are shown as gray spheres ), respectively. SRCR, scavenger receptor cysteine-rich.

Article Snippet: The cDNAs of human SCARA1 to SCARA5 (GenBank: NM_138715.2, NM_006770, NM_016240.2, NM_130386 and NM_286133) were purchased from Sino Biological Inc or synthesized by Sangon Biotech.

Techniques: Binding Assay

Journal: The Journal of Biological Chemistry

Article Title: Recognition of lipoproteins by scavenger receptor class A members

doi: 10.1016/j.jbc.2021.100948

Figure Lengend Snippet: SCARA1, MARCO, and SCARA5 interact with the modified lipoproteins via apoB. A , FACS data for the inhibition of Ac-apoB on the interaction of hSCARA1 with AcLDL. ApoB was applied as a control. B , FACS data for the inhibition of the Ox-apoB or anti-OxPC antibody on the interaction of hSCARA1 with OxLDL. C , FACS data for the inhibition of Ac-apoB on the interaction of hMARCO with AcLDL. D , FACS data for the inhibition of the Ox-apoB or anti-OxPC antibody on the interaction of hMARCO with OxLDL. E , FACS data for the inhibition of Ac-apoB on the interaction of hSCARA5 with AcLDL. F , FACS data for the inhibition of the Ox-apoB or anti-OxPC antibody on the interaction of hSCARA5 with OxLDL. G , ELISA data for the interactions of the SRCR domains of hSCARA1, hMARCO, or hSCARA5 with Ac-apoB or apoB. H , ELISA data for the interactions of the SRCR domains of hSCARA1, hMARCO, or hSCARA5 with Ox-apoB or apoB. I , ELISA data for the interactions of the trimeric CL-SRCR fragment or the monomeric SRCR domain of hSCARA1 with Ac-apoB. J , ELISA data for the interactions of the trimeric CL-SRCR fragment or the monomeric SRCR domain of hSCARA1 with Ox-apoB. ELISA data are representative of three repeated experiments and presented as the mean ± SD. AcLDL, acetylated LDL; CL, collagen-like; OxLDL, oxidized LDL; OxPC, oxidized phosphatidylcholine; SRCR, scavenger receptor cysteine-rich.

Article Snippet: The cDNAs of human SCARA1 to SCARA5 (GenBank: NM_138715.2, NM_006770, NM_016240.2, NM_130386 and NM_286133) were purchased from Sino Biological Inc or synthesized by Sangon Biotech.

Techniques: Modification, Inhibition, Enzyme-linked Immunosorbent Assay

Journal: The Journal of Biological Chemistry

Article Title: Recognition of lipoproteins by scavenger receptor class A members

doi: 10.1016/j.jbc.2021.100948

Figure Lengend Snippet: Interactions of SCARA3 and SCARA4 with lipoproteins and a model for lipoprotein recognition by the SR-A members. A , FACS data of the hSCARA3- or hSCARA4-transfected cells incubated with AcLDL or AcVLDL. Mock represents nontransfected cells. B , FACS data of the hSCARA3- or hSCARA4-transfected cells incubated with OxLDL or OxVLDL. C , ELISA data for the interactions of the CL region of hSCARA3 with AcLDL or OxLDL in the presence of Ca 2+ (2 mM). The SRCR domain of SCARA1 was applied as a control. D , ELISA data for the interactions of the CL region of hSCARA3 with OxVLDL in the presence of Ca 2+ (2 mM). E , FACS data of the hSCARA4-transfected cells incubated with the modified LDL or VLDL in the presence of Ca 2+ or EDTA. F , a cartoon representation of the recognition of the modified LDL/VLDL by the SR-A members SCARA1, MARCO, and SCARA5. ELISA data are representative of three repeated experiments and presented as the mean ± SD. AcLDL, acetylated LDL; AcVLDL, acetylated VLDL; apoB, apolipoprotein B; CL, collagen-like; LDL, low-density lipoprotein; OxLDL, oxidized LDL; OxVLDL, oxidized VLDL; SRCR, scavenger receptor cysteine-rich; VLDL, very-low-density lipoproteins.

Article Snippet: The cDNAs of human SCARA1 to SCARA5 (GenBank: NM_138715.2, NM_006770, NM_016240.2, NM_130386 and NM_286133) were purchased from Sino Biological Inc or synthesized by Sangon Biotech.

Techniques: Transfection, Incubation, Enzyme-linked Immunosorbent Assay, Modification