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cells allophycocyanin apc conjugated mouse monoclonal anti human par  (R&D Systems)


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    R&D Systems cells allophycocyanin apc conjugated mouse monoclonal anti human par
    The cytometric analysis (SSC and FCS) of isolated human resting platelets (A, D) , gated and <t>labeled</t> <t>PAR-1</t> without activation (B, E) , and gated and labeled PAR-1 with activation by 10 µM TRAP (C, F) ; labeled with anti-CD61-FITC and PAR-1-APC antibodies (G–I) . The level of PAR-1 expression was read from gates P1. Markers M1 and M2 indicate the gates for microparticles and normal platelets, with the PAR-1 analysis applied to the summed population. An example image from a patient with DM is shown.
    Cells Allophycocyanin Apc Conjugated Mouse Monoclonal Anti Human Par, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/monoclonal+mouse+anti+human+par1/Human+PAR1+APC-conjugated+Antibody/pmc12856924-121-22-34
    Average 93 stars, based on 13 article reviews
    cells allophycocyanin apc conjugated mouse monoclonal anti human par - by Bioz Stars, 2026-10
    93/100 stars

    Images

    1) Product Images from "The predictive role of protease-activated receptor (PAR-1) polymorphisms and activated microplatelets on the severity of atherosclerosis – preliminary studies"

    Article Title: The predictive role of protease-activated receptor (PAR-1) polymorphisms and activated microplatelets on the severity of atherosclerosis – preliminary studies

    Journal: Frontiers in Molecular Biosciences

    doi: 10.3389/fmolb.2025.1662954

    The cytometric analysis (SSC and FCS) of isolated human resting platelets (A, D) , gated and labeled PAR-1 without activation (B, E) , and gated and labeled PAR-1 with activation by 10 µM TRAP (C, F) ; labeled with anti-CD61-FITC and PAR-1-APC antibodies (G–I) . The level of PAR-1 expression was read from gates P1. Markers M1 and M2 indicate the gates for microparticles and normal platelets, with the PAR-1 analysis applied to the summed population. An example image from a patient with DM is shown.
    Figure Legend Snippet: The cytometric analysis (SSC and FCS) of isolated human resting platelets (A, D) , gated and labeled PAR-1 without activation (B, E) , and gated and labeled PAR-1 with activation by 10 µM TRAP (C, F) ; labeled with anti-CD61-FITC and PAR-1-APC antibodies (G–I) . The level of PAR-1 expression was read from gates P1. Markers M1 and M2 indicate the gates for microparticles and normal platelets, with the PAR-1 analysis applied to the summed population. An example image from a patient with DM is shown.

    Techniques Used: Isolation, Labeling, Activation Assay, Expressing

    The percentage of PAR-1 receptor expression before and after the addition of the thrombin receptor activating peptide (TRAP) in blood samples from patients with diabetic macroangiopathy (DM), the control group (CONTROL), and atherosclerosis obliterans (AO).
    Figure Legend Snippet: The percentage of PAR-1 receptor expression before and after the addition of the thrombin receptor activating peptide (TRAP) in blood samples from patients with diabetic macroangiopathy (DM), the control group (CONTROL), and atherosclerosis obliterans (AO).

    Techniques Used: Expressing, Control

    (A) Separation of DNA molecules in a 3% agarose gel of PAR-1 gene amplification products with the −506 I/D polymorphism. Lanes: 1 – homozygous I/I, 2 – heterozygous I/D, 3 – homozygous D/D, M–GeneRuler™ 50bp DNA Ladder (Fermentas). (B) The percentage distribution of the −506 I/D polymorphism variants in the PAR-1 gene: homozygous D/D (blue), heterozygous I/D (red), and homozygous I/I (green).
    Figure Legend Snippet: (A) Separation of DNA molecules in a 3% agarose gel of PAR-1 gene amplification products with the −506 I/D polymorphism. Lanes: 1 – homozygous I/I, 2 – heterozygous I/D, 3 – homozygous D/D, M–GeneRuler™ 50bp DNA Ladder (Fermentas). (B) The percentage distribution of the −506 I/D polymorphism variants in the PAR-1 gene: homozygous D/D (blue), heterozygous I/D (red), and homozygous I/I (green).

    Techniques Used: Agarose Gel Electrophoresis, Amplification

    (A) The result of the restriction digestion of PCR products with the MvaI enzyme to check for the presence of the −1426 C/T polymorphism in the PAR-1 gene. Lanes: 1 – 6 homozygotes C/C, M–GeneRuler™ 100bp DNA Ladder (Fermentas). (B) The percentage distribution of the variants of the −1426 C/T polymorphism in the PAR-1 gene: homozygote C/C (blue color), heterozygote C/T (red color), homozygote T/T (green color).
    Figure Legend Snippet: (A) The result of the restriction digestion of PCR products with the MvaI enzyme to check for the presence of the −1426 C/T polymorphism in the PAR-1 gene. Lanes: 1 – 6 homozygotes C/C, M–GeneRuler™ 100bp DNA Ladder (Fermentas). (B) The percentage distribution of the variants of the −1426 C/T polymorphism in the PAR-1 gene: homozygote C/C (blue color), heterozygote C/T (red color), homozygote T/T (green color).

    Techniques Used:

    (A) Example separations of amplification products of the DNA fragment encompassing the IVSn-14 A/T polymorphism site of the PAR-1 gene using the SNaPshot method. Alleles were determined based on the size of primers and the colors of fluorescently labeled ddNTPs (terminators) incorporated during the primer extension reaction. (A) red peak, wild-type homozygote (AA); (B) green and red peaks, heterozygote (AT); (C) green peak, mutated homozygote (TT). (B) The percentage distribution of the variants of the IVS-14 A/T polymorphism of the PAR-1 gene is as follows: homozygote A/A (blue color), heterozygote A/T (red color), and homozygote T/T (green color).
    Figure Legend Snippet: (A) Example separations of amplification products of the DNA fragment encompassing the IVSn-14 A/T polymorphism site of the PAR-1 gene using the SNaPshot method. Alleles were determined based on the size of primers and the colors of fluorescently labeled ddNTPs (terminators) incorporated during the primer extension reaction. (A) red peak, wild-type homozygote (AA); (B) green and red peaks, heterozygote (AT); (C) green peak, mutated homozygote (TT). (B) The percentage distribution of the variants of the IVS-14 A/T polymorphism of the PAR-1 gene is as follows: homozygote A/A (blue color), heterozygote A/T (red color), and homozygote T/T (green color).

    Techniques Used: Amplification, Labeling

    Multivariate analysis: (A–D) Number of microparticles with PAR-1+TRAP; (E–F) Number of microparticles with BMI (Figure 4.12E-F) and age with smoking.
    Figure Legend Snippet: Multivariate analysis: (A–D) Number of microparticles with PAR-1+TRAP; (E–F) Number of microparticles with BMI (Figure 4.12E-F) and age with smoking.

    Techniques Used:

    Related Articles

    Solvent:

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro.
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Recombinant:

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro.
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    SDS-Gel:

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro.
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Electrophoresis:

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro.
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Western Blot:

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro.
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Staining:

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro.
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Comparison:

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro.
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Expressing:

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro.
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Control:

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro.
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Fluorescence:

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro.
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Incubation:

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro.
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Microscopy:

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro.
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Software:

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro.
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Transfection:

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro.
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro
    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).



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    cells allophycocyanin apc conjugated mouse monoclonal anti human par  (R&D Systems)
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    R&D Systems cells allophycocyanin apc conjugated mouse monoclonal anti human par
    The cytometric analysis (SSC and FCS) of isolated human resting platelets (A, D) , gated and <t>labeled</t> <t>PAR-1</t> without activation (B, E) , and gated and labeled PAR-1 with activation by 10 µM TRAP (C, F) ; labeled with anti-CD61-FITC and PAR-1-APC antibodies (G–I) . The level of PAR-1 expression was read from gates P1. Markers M1 and M2 indicate the gates for microparticles and normal platelets, with the PAR-1 analysis applied to the summed population. An example image from a patient with DM is shown.
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    The cytometric analysis (SSC and FCS) of isolated human resting platelets (A, D) , gated and <t>labeled</t> <t>PAR-1</t> without activation (B, E) , and gated and labeled PAR-1 with activation by 10 µM TRAP (C, F) ; labeled with anti-CD61-FITC and PAR-1-APC antibodies (G–I) . The level of PAR-1 expression was read from gates P1. Markers M1 and M2 indicate the gates for microparticles and normal platelets, with the PAR-1 analysis applied to the summed population. An example image from a patient with DM is shown.
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    Figure 4: MMP1 induces Ca2+ signalling, and activates migratory proteins and CCIDs in LECs. a. LECs were grown to ~70–80% confluence and then pre-treated with 0.5 μM SCH79797 or solvent (DMSO) and then stimulated with 100 ng/ml activated recombinant MMP1 for 4 h. Cells were lysed, proteins were separated by SDS gel electrophoresis and analysed by Western blotting using the indicated antibodies. Equal sample loading was controlled by Ponceau S staining and ß-actin immunoblotting. Densitometer readings facilitated the comparison of relative protein expression levels with solvent treated control (which was set as “1”). (b) LECs (8 × 103 cells/ well) were pre-treated with 0.5 μM SCH79797 <t>(PAR1</t> inhibitor) and then charged with FluoForte Dye-loading in presence of SCH79797 for 45 min at 37°C and 15 min at room temperature. Then, cells were stimulated with 100 ng/ml activated recombinant MMP1 for 5 min. Intracellular free calcium was measured with a fluorescence plate reader at 490/525 nm. Experiments were performed in triplicate, error bars indicate means +/− SEM, and asterisks and rhomboids significance (p < 0.05; t-test). (c) Confluent LECs were pre-treated with SCH79797 or solvent (DMSO) for 30 min and then MDA-MB231 spheroids were placed on top of LECs monolayers and co-incubated for 4 h. The areas of CCIDs were analysed using an Axiovert microscope and Zen Little 2012 software.
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    Image Search Results


    The cytometric analysis (SSC and FCS) of isolated human resting platelets (A, D) , gated and labeled PAR-1 without activation (B, E) , and gated and labeled PAR-1 with activation by 10 µM TRAP (C, F) ; labeled with anti-CD61-FITC and PAR-1-APC antibodies (G–I) . The level of PAR-1 expression was read from gates P1. Markers M1 and M2 indicate the gates for microparticles and normal platelets, with the PAR-1 analysis applied to the summed population. An example image from a patient with DM is shown.

    Journal: Frontiers in Molecular Biosciences

    Article Title: The predictive role of protease-activated receptor (PAR-1) polymorphisms and activated microplatelets on the severity of atherosclerosis – preliminary studies

    doi: 10.3389/fmolb.2025.1662954

    Figure Lengend Snippet: The cytometric analysis (SSC and FCS) of isolated human resting platelets (A, D) , gated and labeled PAR-1 without activation (B, E) , and gated and labeled PAR-1 with activation by 10 µM TRAP (C, F) ; labeled with anti-CD61-FITC and PAR-1-APC antibodies (G–I) . The level of PAR-1 expression was read from gates P1. Markers M1 and M2 indicate the gates for microparticles and normal platelets, with the PAR-1 analysis applied to the summed population. An example image from a patient with DM is shown.

    Article Snippet: Next: For the PAR-1 test without PLT activation, 5 μL of PAR-1-APC antibodies at a concentration of 5 μg/5 × 10 5 cells (Allophycocyanin (APC)-conjugated mouse monoclonal anti-human PAR-1; clone# 731115; mouse isotype: IgG1, R&D Systems, Minneapolis, Canada) and 5 μL of CD61-FITC antibodies (Monoclonal Mouse Anti-Human CD61, Platelet Glycoprotein IIIa/FITC, Clone Y2/51, code: F0803, DakoCytomation, Glostrup, Denmark) were added.

    Techniques: Isolation, Labeling, Activation Assay, Expressing

    The percentage of PAR-1 receptor expression before and after the addition of the thrombin receptor activating peptide (TRAP) in blood samples from patients with diabetic macroangiopathy (DM), the control group (CONTROL), and atherosclerosis obliterans (AO).

    Journal: Frontiers in Molecular Biosciences

    Article Title: The predictive role of protease-activated receptor (PAR-1) polymorphisms and activated microplatelets on the severity of atherosclerosis – preliminary studies

    doi: 10.3389/fmolb.2025.1662954

    Figure Lengend Snippet: The percentage of PAR-1 receptor expression before and after the addition of the thrombin receptor activating peptide (TRAP) in blood samples from patients with diabetic macroangiopathy (DM), the control group (CONTROL), and atherosclerosis obliterans (AO).

    Article Snippet: Next: For the PAR-1 test without PLT activation, 5 μL of PAR-1-APC antibodies at a concentration of 5 μg/5 × 10 5 cells (Allophycocyanin (APC)-conjugated mouse monoclonal anti-human PAR-1; clone# 731115; mouse isotype: IgG1, R&D Systems, Minneapolis, Canada) and 5 μL of CD61-FITC antibodies (Monoclonal Mouse Anti-Human CD61, Platelet Glycoprotein IIIa/FITC, Clone Y2/51, code: F0803, DakoCytomation, Glostrup, Denmark) were added.

    Techniques: Expressing, Control

    (A) Separation of DNA molecules in a 3% agarose gel of PAR-1 gene amplification products with the −506 I/D polymorphism. Lanes: 1 – homozygous I/I, 2 – heterozygous I/D, 3 – homozygous D/D, M–GeneRuler™ 50bp DNA Ladder (Fermentas). (B) The percentage distribution of the −506 I/D polymorphism variants in the PAR-1 gene: homozygous D/D (blue), heterozygous I/D (red), and homozygous I/I (green).

    Journal: Frontiers in Molecular Biosciences

    Article Title: The predictive role of protease-activated receptor (PAR-1) polymorphisms and activated microplatelets on the severity of atherosclerosis – preliminary studies

    doi: 10.3389/fmolb.2025.1662954

    Figure Lengend Snippet: (A) Separation of DNA molecules in a 3% agarose gel of PAR-1 gene amplification products with the −506 I/D polymorphism. Lanes: 1 – homozygous I/I, 2 – heterozygous I/D, 3 – homozygous D/D, M–GeneRuler™ 50bp DNA Ladder (Fermentas). (B) The percentage distribution of the −506 I/D polymorphism variants in the PAR-1 gene: homozygous D/D (blue), heterozygous I/D (red), and homozygous I/I (green).

    Article Snippet: Next: For the PAR-1 test without PLT activation, 5 μL of PAR-1-APC antibodies at a concentration of 5 μg/5 × 10 5 cells (Allophycocyanin (APC)-conjugated mouse monoclonal anti-human PAR-1; clone# 731115; mouse isotype: IgG1, R&D Systems, Minneapolis, Canada) and 5 μL of CD61-FITC antibodies (Monoclonal Mouse Anti-Human CD61, Platelet Glycoprotein IIIa/FITC, Clone Y2/51, code: F0803, DakoCytomation, Glostrup, Denmark) were added.

    Techniques: Agarose Gel Electrophoresis, Amplification

    (A) The result of the restriction digestion of PCR products with the MvaI enzyme to check for the presence of the −1426 C/T polymorphism in the PAR-1 gene. Lanes: 1 – 6 homozygotes C/C, M–GeneRuler™ 100bp DNA Ladder (Fermentas). (B) The percentage distribution of the variants of the −1426 C/T polymorphism in the PAR-1 gene: homozygote C/C (blue color), heterozygote C/T (red color), homozygote T/T (green color).

    Journal: Frontiers in Molecular Biosciences

    Article Title: The predictive role of protease-activated receptor (PAR-1) polymorphisms and activated microplatelets on the severity of atherosclerosis – preliminary studies

    doi: 10.3389/fmolb.2025.1662954

    Figure Lengend Snippet: (A) The result of the restriction digestion of PCR products with the MvaI enzyme to check for the presence of the −1426 C/T polymorphism in the PAR-1 gene. Lanes: 1 – 6 homozygotes C/C, M–GeneRuler™ 100bp DNA Ladder (Fermentas). (B) The percentage distribution of the variants of the −1426 C/T polymorphism in the PAR-1 gene: homozygote C/C (blue color), heterozygote C/T (red color), homozygote T/T (green color).

    Article Snippet: Next: For the PAR-1 test without PLT activation, 5 μL of PAR-1-APC antibodies at a concentration of 5 μg/5 × 10 5 cells (Allophycocyanin (APC)-conjugated mouse monoclonal anti-human PAR-1; clone# 731115; mouse isotype: IgG1, R&D Systems, Minneapolis, Canada) and 5 μL of CD61-FITC antibodies (Monoclonal Mouse Anti-Human CD61, Platelet Glycoprotein IIIa/FITC, Clone Y2/51, code: F0803, DakoCytomation, Glostrup, Denmark) were added.

    Techniques:

    (A) Example separations of amplification products of the DNA fragment encompassing the IVSn-14 A/T polymorphism site of the PAR-1 gene using the SNaPshot method. Alleles were determined based on the size of primers and the colors of fluorescently labeled ddNTPs (terminators) incorporated during the primer extension reaction. (A) red peak, wild-type homozygote (AA); (B) green and red peaks, heterozygote (AT); (C) green peak, mutated homozygote (TT). (B) The percentage distribution of the variants of the IVS-14 A/T polymorphism of the PAR-1 gene is as follows: homozygote A/A (blue color), heterozygote A/T (red color), and homozygote T/T (green color).

    Journal: Frontiers in Molecular Biosciences

    Article Title: The predictive role of protease-activated receptor (PAR-1) polymorphisms and activated microplatelets on the severity of atherosclerosis – preliminary studies

    doi: 10.3389/fmolb.2025.1662954

    Figure Lengend Snippet: (A) Example separations of amplification products of the DNA fragment encompassing the IVSn-14 A/T polymorphism site of the PAR-1 gene using the SNaPshot method. Alleles were determined based on the size of primers and the colors of fluorescently labeled ddNTPs (terminators) incorporated during the primer extension reaction. (A) red peak, wild-type homozygote (AA); (B) green and red peaks, heterozygote (AT); (C) green peak, mutated homozygote (TT). (B) The percentage distribution of the variants of the IVS-14 A/T polymorphism of the PAR-1 gene is as follows: homozygote A/A (blue color), heterozygote A/T (red color), and homozygote T/T (green color).

    Article Snippet: Next: For the PAR-1 test without PLT activation, 5 μL of PAR-1-APC antibodies at a concentration of 5 μg/5 × 10 5 cells (Allophycocyanin (APC)-conjugated mouse monoclonal anti-human PAR-1; clone# 731115; mouse isotype: IgG1, R&D Systems, Minneapolis, Canada) and 5 μL of CD61-FITC antibodies (Monoclonal Mouse Anti-Human CD61, Platelet Glycoprotein IIIa/FITC, Clone Y2/51, code: F0803, DakoCytomation, Glostrup, Denmark) were added.

    Techniques: Amplification, Labeling

    Multivariate analysis: (A–D) Number of microparticles with PAR-1+TRAP; (E–F) Number of microparticles with BMI (Figure 4.12E-F) and age with smoking.

    Journal: Frontiers in Molecular Biosciences

    Article Title: The predictive role of protease-activated receptor (PAR-1) polymorphisms and activated microplatelets on the severity of atherosclerosis – preliminary studies

    doi: 10.3389/fmolb.2025.1662954

    Figure Lengend Snippet: Multivariate analysis: (A–D) Number of microparticles with PAR-1+TRAP; (E–F) Number of microparticles with BMI (Figure 4.12E-F) and age with smoking.

    Article Snippet: Next: For the PAR-1 test without PLT activation, 5 μL of PAR-1-APC antibodies at a concentration of 5 μg/5 × 10 5 cells (Allophycocyanin (APC)-conjugated mouse monoclonal anti-human PAR-1; clone# 731115; mouse isotype: IgG1, R&D Systems, Minneapolis, Canada) and 5 μL of CD61-FITC antibodies (Monoclonal Mouse Anti-Human CD61, Platelet Glycoprotein IIIa/FITC, Clone Y2/51, code: F0803, DakoCytomation, Glostrup, Denmark) were added.

    Techniques:

    Figure 4: MMP1 induces Ca2+ signalling, and activates migratory proteins and CCIDs in LECs. a. LECs were grown to ~70–80% confluence and then pre-treated with 0.5 μM SCH79797 or solvent (DMSO) and then stimulated with 100 ng/ml activated recombinant MMP1 for 4 h. Cells were lysed, proteins were separated by SDS gel electrophoresis and analysed by Western blotting using the indicated antibodies. Equal sample loading was controlled by Ponceau S staining and ß-actin immunoblotting. Densitometer readings facilitated the comparison of relative protein expression levels with solvent treated control (which was set as “1”). (b) LECs (8 × 103 cells/ well) were pre-treated with 0.5 μM SCH79797 (PAR1 inhibitor) and then charged with FluoForte Dye-loading in presence of SCH79797 for 45 min at 37°C and 15 min at room temperature. Then, cells were stimulated with 100 ng/ml activated recombinant MMP1 for 5 min. Intracellular free calcium was measured with a fluorescence plate reader at 490/525 nm. Experiments were performed in triplicate, error bars indicate means +/− SEM, and asterisks and rhomboids significance (p < 0.05; t-test). (c) Confluent LECs were pre-treated with SCH79797 or solvent (DMSO) for 30 min and then MDA-MB231 spheroids were placed on top of LECs monolayers and co-incubated for 4 h. The areas of CCIDs were analysed using an Axiovert microscope and Zen Little 2012 software.

    Journal: Oncotarget

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro.

    doi: 10.18632/oncotarget.5741

    Figure Lengend Snippet: Figure 4: MMP1 induces Ca2+ signalling, and activates migratory proteins and CCIDs in LECs. a. LECs were grown to ~70–80% confluence and then pre-treated with 0.5 μM SCH79797 or solvent (DMSO) and then stimulated with 100 ng/ml activated recombinant MMP1 for 4 h. Cells were lysed, proteins were separated by SDS gel electrophoresis and analysed by Western blotting using the indicated antibodies. Equal sample loading was controlled by Ponceau S staining and ß-actin immunoblotting. Densitometer readings facilitated the comparison of relative protein expression levels with solvent treated control (which was set as “1”). (b) LECs (8 × 103 cells/ well) were pre-treated with 0.5 μM SCH79797 (PAR1 inhibitor) and then charged with FluoForte Dye-loading in presence of SCH79797 for 45 min at 37°C and 15 min at room temperature. Then, cells were stimulated with 100 ng/ml activated recombinant MMP1 for 5 min. Intracellular free calcium was measured with a fluorescence plate reader at 490/525 nm. Experiments were performed in triplicate, error bars indicate means +/− SEM, and asterisks and rhomboids significance (p < 0.05; t-test). (c) Confluent LECs were pre-treated with SCH79797 or solvent (DMSO) for 30 min and then MDA-MB231 spheroids were placed on top of LECs monolayers and co-incubated for 4 h. The areas of CCIDs were analysed using an Axiovert microscope and Zen Little 2012 software.

    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Techniques: Solvent, Recombinant, SDS-Gel, Electrophoresis, Western Blot, Staining, Comparison, Expressing, Control, Fluorescence, Incubation, Microscopy, Software

    Figure 5: Combined inhibitions of NF-κB and target genes in MDA-MB231 spheroids and LEC monolayers. LECs or MDA-MB231 spheroids were transiently transfected with either non-targeting (n.t.) siRNA or a combination of siRNAs inhibiting the expression of a. NFKB1 & NFKB2, b. or siRNAs inhibiting the expression of ICAM-1 and MMP1 respectively. After 24 h, spheroids were placed on top of confluent LEC monolayers and co-incubated for 4 h. c. MDA-MB231 spheroids were transiently transfected with either non-targeting (n.t.) siRNA or siRNA inhibiting the expression of MMP1. After 24 h, spheroids were placed on top of confluent LEC monolayers, which were pre-treated with 1 μM SCH79797 (inhibitor of PAR1) or solvent (DMSO) for 30 min, and co-incubated for 4 h. The areas of CCIDs were analysed using an Axiovert microscope and Zen Little 2012 software. Experiments were performed in triplicate, error bars indicate means +/− SEM, and asterisks and rhomboids significance (p < 0.05; t-test).

    Journal: Oncotarget

    Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro.

    doi: 10.18632/oncotarget.5741

    Figure Lengend Snippet: Figure 5: Combined inhibitions of NF-κB and target genes in MDA-MB231 spheroids and LEC monolayers. LECs or MDA-MB231 spheroids were transiently transfected with either non-targeting (n.t.) siRNA or a combination of siRNAs inhibiting the expression of a. NFKB1 & NFKB2, b. or siRNAs inhibiting the expression of ICAM-1 and MMP1 respectively. After 24 h, spheroids were placed on top of confluent LEC monolayers and co-incubated for 4 h. c. MDA-MB231 spheroids were transiently transfected with either non-targeting (n.t.) siRNA or siRNA inhibiting the expression of MMP1. After 24 h, spheroids were placed on top of confluent LEC monolayers, which were pre-treated with 1 μM SCH79797 (inhibitor of PAR1) or solvent (DMSO) for 30 min, and co-incubated for 4 h. The areas of CCIDs were analysed using an Axiovert microscope and Zen Little 2012 software. Experiments were performed in triplicate, error bars indicate means +/− SEM, and asterisks and rhomboids significance (p < 0.05; t-test).

    Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

    Techniques: Transfection, Expressing, Incubation, Solvent, Microscopy, Software