Journal: Cell Reports Medicine

Article Title: Spleen-targeted neoantigen mRNA vaccine induces ISG15 + CD8 + T cell-mediated tertiary lymphoid structure formation in hepatocellular carcinoma

doi: 10.1016/j.xcrm.2026.102754

Figure Lengend Snippet: GZMA-F2R interaction mediates STNvac efficacy through T cell activation (A) Bubble plot showing ligand-receptor pairs between ISG15 + CD8 + T cells and APCs (B cells, CD4 + T cells, and DCs). (B) Violin plots showing F2R expression across T cell subsets and GZMA expression in APCs. (C and D) Therapeutic evaluation of STNvac co-administration with F2R antagonist (F2RA, SCH79797 ) ( n = 7 mice per group). (C) Bioluminescence images showing tumor burden in orthotopic HCC-bearing mice under the indicated treatments. (D) Survival curves of mice in different groups. (E and F) Multicolor immunofluorescence staining of intratumoral Ki67 + CD69 + ISG15 + CD8 + T cells. (E) Representative images. Scale bars, 20 μm. (F) Quantitative analysis of positive cell density in five randomly selected areas per tumor section. (G and H) Activation of human HCC TILs through the GZMA-F2R interaction. (G) Schematic illustration of the treatment schedule. (H) Quantitative analysis of 41BB + CD3 + CD8 + T cells after 24 h of GZMA stimulation ( n = 2 biological replicates, each analyzed in triplicate). Statistics: one-way ANOVA for (F) and (H); log rank (Mantel-Cox) test for (D). Mean ± SD. Significance levels: ∗p < 0.05, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. See also and .

Article Snippet: To evaluate the impact of the GZMA-F2R signaling on CD8 + TIL activation, the in vitro-expanded TILs were seeded in 24-well plates, pre-treated with the F2R antagonist SCH79797 (MCE, 5 μM), and subsequently stimulated with recombinant GZMA (MCE, 0.05 ng/mL).

Techniques: Activation Assay, Expressing, Multicolor Immunofluorescence Staining

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

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

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

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:

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

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: