Journal: JCI Insight

Article Title: Platelets impair the resolution of inflammation in atherosclerotic plaques in insulin-resistant mice after lipid lowering

doi: 10.1172/jci.insight.193593

Figure Lengend Snippet: ( A ) Study design. Eight-week-old Ldlr –/– male mice fed a high-fat, high-cholesterol (HFHC) diet for the whole duration of the study (23 weeks). After the atherosclerosis progression period (obese baseline, Ob), mice were injected with ApoB ASO to lower lipids (lipid-lowering period). All lipid-lowered mice were split into 2 groups, injected either with isotype control antibody (IgG) or platelet depletion antibody (αCD42b) every 3 days for a total of 3 weeks. ( B ) Platelet counts in circulating blood of Ob, IgG, and αCD42b 3 days before harvest. ( C ) Blood glucose measurements (lean n = 16, Ob n = 47) and ( D ) area of curve (AOC) quantification of glucose tolerance tests (GTTs) from lean, Ob, IgG, and αCD42b mice after 16 weeks of HFHC diet during atherosclerosis progression. ( E ) Blood glucose measurements (Ob n = 3, IgG n = 4, αCD42b n = 4) and ( F ) AOC quantification of GTTs from Ob, IgG, and αCD42b mice after 22 weeks of HFHC diet during atherosclerosis resolution. Mean fluorescent intensity (MFI) of platelet ( G ) JonA and ( H ) P-selectin in nonstimulated (NS) samples and upon 100 μM PAR4-AP agonist stimulation of lean and Ob circulating blood samples. ( I ) Mean platelet volume (MPV) from circulating blood of lean, Ob, and IgG-treated mice, 3 days before harvest. In C and E , error bars represent SD. Data were analyzed by Kruskal-Wallis with Dunn’s post hoc test ( B and F ), unpaired Mann-Whitney test ( C , D , and H ), unpaired Student’s t test ( G ), or ordinary 1-way ANOVA with Tukey’s multiple-comparison test ( I ). P values are shown in graphs.

Article Snippet: The whole blood was stained with anti–mouse JonA-PE (M023-2, Emfret Analytics) and anti–mouse P-selectin Alexa Fluor 647 (563674, BD Biosciences) and then treated with PAR4-AP platelet agonist (100 μM; MedChemExpress) for 15 minutes at room temperature.

Techniques: Injection, Control, MANN-WHITNEY, Comparison

Journal: Frontiers in Pharmacology

Article Title: Role of albumin in regulating platelet function

doi: 10.3389/fphar.2026.1734694

Figure Lengend Snippet: Hypoproteinemia was found to promote platelet activation. (A–E) Wash platelet aggregation in WT, ADR and ADR Albumin mice induced by thrombin (A) , U46619 (B) , PAR4-AP (C) , collagen (D) and ADP (E) (n = 5 independent experimental animals). (F–H) Thrombin (F) , U46619 (G) and PAR4-AP (H) induced integrin αIIbβ3 activation in washed platelets from mice (n = 5 independent experimental animals). (I–K) Thrombin (I) , U46619 (J) and PAR4-AP (K) induced P-selectin release from washed platelets from mice (n = 5 independent experimental animals). (L–O) ATP release from washed platelets in mice was induced by thrombin (L) , U46619 (M) , PAR4-AP (N) and collagen (O) (n = 5 independent experimental animals). (P–R) Wash platelet spreading in mice induced by U46619 (P) , mean spreading area of individual platelets (Q) , and proportion of spreading platelets to total platelets in the field of view (R) (n = 3 independent experimental animals). Differences between groups were assessed by one-way ANOVA followed by Dunnett’s post hoc test. Statistics are presented as mean ± SEM. * P < 0.05, ** P < 0.01 and *** P < 0.001.

Article Snippet: Rabbit antibodies against phospho-PKC substrate (2261), phospho-Akt (Ser473) (11E7), Akt (pan) (11E7), PKC alpha (2056), GAPDH (5174), cell lysate buffer (9803), and PAR4-AP (2328) were sourced from Cell Signaling Technology, Inc. (Beverly, MA, United States).

Techniques: Activation Assay

Journal: Frontiers in Oncology

Article Title: The protease-activated receptors are expressed in glioblastoma and differentially modulate adherent versus stem-like growth of LN-18 GBM cells

doi: 10.3389/fonc.2025.1582996

Figure Lengend Snippet: PAR1–4 mRNA expression in GBM specimen and its association with patients´ overall survival time. (A) Comparison of PAR1, PAR2, PAR3 and PAR4 mRNA expression in non-malignant brain (NMB, n=7) and all analyzed GBM samples (both primary and relapsed GBM, n=118). (B) Subdivision of GBM specimen in primary GBM (prGBM, n=78), first (1 st , n=33) and second (2 nd , n=7) relapses and comparison with NMB. (A+B) Gene expression was measured by qPCR. Each mRNA level of the target genes (PAR1-4) was normalized to the mean of GAPDH and β-actin using the 2 -ΔΔct method. Data are shown as scatter plots representing the median as horizontal bars. Mann Whitney U test, *p < 0.05 and ***p < 0.001 for (A) and OneWay ANOVA/Kruskal Wallis test with Dunn’s Multiple Comparison Test, *p < 0.05, **p < 0.01 and *** p < 0.001 for (B) . (C) Kaplan Meier survival analyses of PAR1–4 mRNA expression in GBM patients. Association of the relative mRNA expression of each single PAR receptor with the survival time of patients with primary GBM. The patients were divided into two subgroups depending on the median gene expression. No significant association was found.

Article Snippet: As specific PAR antagonists the following compounds were used: PAR1 antagonist RWJ, PAR2 antagonist FSLLRY-NH2, and PAR4 antagonist tcY-NH2 (all from Tocris Bioscience).

Techniques: Expressing, Comparison, Gene Expression, MANN-WHITNEY

Journal: Frontiers in Oncology

Article Title: The protease-activated receptors are expressed in glioblastoma and differentially modulate adherent versus stem-like growth of LN-18 GBM cells

doi: 10.3389/fonc.2025.1582996

Figure Lengend Snippet: Impact of PAR subtype specific inhibitors and agonists on viability of LN-18 GBM cells. Adherent and neurospheric LN-18 cells of passage 1 or 3 were treated for 48 or 72h with the respective compounds followed by measurement of cell viability using the Resazurin assay. (A+B) Treatment of adherent (A) and neurospheric (B) LN-18 cells with inhibitors of PAR1 (RWJ), PAR2 (FSLLRY) and PAR4 (tcY) (each 5 and 20 µM), n=3-4. (C+D) Incubation of adherent (C) and neurospheric (D) LN-18 cells with Thrombin (TB, 30 U/ml), Factor Xa (FXa, 30 nM) or PAR subtype specific agonists (AP1 to AP4, each 100 and 200 µM), n=3-4. OneWay ANOVA Dunnett’s Multiple Comparison Test, *p < 0.05, **p < 0.01 and ***p < 0.001 vs. Con.

Article Snippet: As specific PAR antagonists the following compounds were used: PAR1 antagonist RWJ, PAR2 antagonist FSLLRY-NH2, and PAR4 antagonist tcY-NH2 (all from Tocris Bioscience).

Techniques: Resazurin Assay, Incubation, Comparison