par1 activating peptide  (Cosmo Genetech Co)

Journal: International Journal of Molecular Sciences

Article Title: Blockade of PAR2 Signaling by Punicalagin as a Therapeutic Strategy for Atopic Dermatitis

doi: 10.3390/ijms26188920

Figure Lengend Snippet: Potency and selectivity of PCG for PAR2. ( A , B ) Inhibition of PAR2-mediated calcium mobilization by PCG in HaCaT cells. Cells were pretreated with PCG for 10 min before stimulation with 30 µM PAR2-AP or 30 U/mL trypsin (TRY). ( C , D ) Effect of PCG on PAR1-mediated calcium mobilization in HaCaT cells. Cells were pretreated with PCG for 10 min before stimulation with 30 µM PAR1-AP or 30 U/mL thrombin (TRB). ( E ) Summary of dose–response curves for PAR2 and PAR1 inhibition by PCG.

Article Snippet: PAR1 activating peptide (PAR1-AP, TRLLR-NH 2 ) and PAR2 activating peptide (PAR2-AP, SLIGRL-NH 2 ) were synthesized by Cosmogenetech Co., Ltd. (Seoul, Republic of Korea).

Techniques: Inhibition

Journal: TH Open: Companion Journal to Thrombosis and Haemostasis

Article Title: Platelet Activation Pathways Controlling Reversible Integrin αIIbβ3 Activation

doi: 10.1055/s-0044-1786987

Figure Lengend Snippet: Time-dependent effects of GPVI- and PAR-induced integrin αIIbβ3 activation and P-selectin expression. Washed platelets were stimulated for 10 or 20 minutes with indicated doses of CRP (μg/mL), thrombin (nM), TRAP6 (μM) or AYPGKF (AY, μM). The platelet samples were then labeled with FITC-PAC1 mAb and AF647 anti-CD62P mAb for active intgrins and P-selectin expression, respectively, for analysis by flow cytometry. Shown are percentages of positive platelets in response to CRP ( A ), thrombin ( B ), TRAP6 ( C ), or AYPGFK ( D ). Representative histograms are shown in Suppl. Figure 1. Data are means ± SD ( n = 3–6); * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 for 10 min vs. 20 min ( t -test).

Article Snippet: The platelet agonist thrombin receptor-activating peptide 6, TRAP6 (SFLLRN, PAR1), was purchased from Bachem (Bubendorf, Switzerland); cross-linked CRP was obtained from CambCol (Cambridge, United Kingdom); the PAR4 agonist AYPGKF was from Bio-Techne (Minneapolis, Minnesota, United States).

Techniques: Activation Assay, Expressing, Labeling, Flow Cytometry

Journal: TH Open: Companion Journal to Thrombosis and Haemostasis

Article Title: Platelet Activation Pathways Controlling Reversible Integrin αIIbβ3 Activation

doi: 10.1055/s-0044-1786987

Figure Lengend Snippet: Effects of signaling inhibitors on GPVI- and PAR-induced αIIbβ3 activation and P-selectin expression. Washed platelets were pretreated for 10 minutes with vehicle (none) or optimized doses of Gö6976 (1 µM), ML-099 (10 µM), PKCθ-IN (2.5 µM), TGX-221 (0.5 µM), DM-BAPTA (50 µM, 30 minutes), rottlerin (10 µM), ML-314 (20 µM), GSK3-IN (2 µM) or RO-318425 (10 µM). Subsequently, the platelet samples were left unstimulated or stimulated with CRP (5 or 2.5 µg/mL), TRAP6 (15 or 7.5 µM), thrombin (5 or 2.5 nM) or AYPGKF (50 µM) for 10 mins. Using flow cytometry, αIIbβ3 activation (% PAC1 positive) and P-selectin expression (% CD62P positive) were assessed, as in Figure 1. Data were scaled 0-10 from unstimulated to maximal activation marker, based on mean values ( n = 3-6). ( A ) Unsupervised clustering of scaled values of αIIbβ3 activation ( i ), and percentage change by indicated signaling inhibitor ( ii ). The resulting ordering of agonists and inhibitors was used for all heatmap presentations. ( B ) Corresponding heatmaps of extent of P-selectin expression ( i ) and differential percentage change by indicated signaling inhibitor ( ii ). For the posttreatment condition, platelets were left unstimulated or stimulated for 10 minutes with CRP (5 or 2.5 µg/mL), TRAP6 (15 or 7.5 µM), thrombin (5 or 2.5 nM) or AYPGKF (50 µM). After 10 minutes, the cells were posttreated with vehicle (none) or indicated signaling inhibitors, as mentioned above. After 10 minutes, integrin αIIbβ3 activation (% PAC1 positive) and P-selectin expression (% CD62P positive) were measured by flow cytometry. Shown are heatmaps of procentual changes in positive platelets versus no inhibitor for αIIbβ3 activation ( C ) or P-selectin expression ( D ). Color bars refer to scaling used. Means ( n ≥ 3); * p < 0.05, ** p < 0.01, *** p < 0.001 for inhibitor vs. control condition (none) without inhibitor at endpoint ( t -test).

Article Snippet: The platelet agonist thrombin receptor-activating peptide 6, TRAP6 (SFLLRN, PAR1), was purchased from Bachem (Bubendorf, Switzerland); cross-linked CRP was obtained from CambCol (Cambridge, United Kingdom); the PAR4 agonist AYPGKF was from Bio-Techne (Minneapolis, Minnesota, United States).

Techniques: Activation Assay, Expressing, Flow Cytometry, Marker, Control

Journal: TH Open: Companion Journal to Thrombosis and Haemostasis

Article Title: Platelet Activation Pathways Controlling Reversible Integrin αIIbβ3 Activation

doi: 10.1055/s-0044-1786987

Figure Lengend Snippet: Time effects of ADP receptor-dependent integrin αIIbβ3 activation and P-selectin expression. Platelets were stimulated for 10 or 20 minutes as indicated with CRP (5 μg/mL), TRAP6 (15 μM) and/or ADP (1 μM). Samples were pre-incubated with ADP-degrading apyrase (1 U/mL). Integrin αIIbβ3 activation ( i ) and P-selectin expression ( ii ) were measured, as for . Shown are percentage of positive platelets in response to CRP and/or ADP ( A ), or to TRAP6 and/or ADP ( B ), with additional quantification ( C ). Means ± SD ( n ≥ 3); * p < 0.05, ** p < 0.01, **** p < 0.0001 for 10 min vs. 20 min ( t -test). Representative histograms in .

Article Snippet: The platelet agonist thrombin receptor-activating peptide 6, TRAP6 (SFLLRN, PAR1), was purchased from Bachem (Bubendorf, Switzerland); cross-linked CRP was obtained from CambCol (Cambridge, United Kingdom); the PAR4 agonist AYPGKF was from Bio-Techne (Minneapolis, Minnesota, United States).

Techniques: Activation Assay, Expressing, Incubation

Journal: TH Open: Companion Journal to Thrombosis and Haemostasis

Article Title: Platelet Activation Pathways Controlling Reversible Integrin αIIbβ3 Activation

doi: 10.1055/s-0044-1786987

Figure Lengend Snippet: Role of ADP on GPVI- and PAR-induced integrin αIIbβ3 activation and P-selectin expression. In the pretreatment condition ( A ), platelets were incubated for 10 minutes with vehicle (none) or optimized doses of ticagrelor (1 µM) and/or MRS-2179 (50 µM). The cells were subsequently stimulated with CRP (5 or 2.5 µg/mL), TRAP6 (15 or 7.5 µM), thrombin (5 or 2.5 nM) or AYPGKF (50 µM). In the posttreatment condition ( B ), the platelets were left unstimulated or stimulated with the same agonists. After 10 minutes, the platelets were posttreated with vehicle (none) or ticagrelor and/or MRS-2179. Flow cytometric detection was performed 10 minutes later, as in . Shown are heatmaps of extent of αIIbβ3 activation ( i ), and P-selectin expression ( ii ). Means ( n = 3–4); * p < 0.05, ** p < 0.01, *** p < 0.001 for inhibitor vs. control condition (none) without inhibitor at endpoint ( t -test). For graphs of raw data (PAC positive, % of control), see .

Article Snippet: The platelet agonist thrombin receptor-activating peptide 6, TRAP6 (SFLLRN, PAR1), was purchased from Bachem (Bubendorf, Switzerland); cross-linked CRP was obtained from CambCol (Cambridge, United Kingdom); the PAR4 agonist AYPGKF was from Bio-Techne (Minneapolis, Minnesota, United States).

Techniques: Activation Assay, Expressing, Incubation, Control, Positive Control

Journal: TH Open: Companion Journal to Thrombosis and Haemostasis

Article Title: Platelet Activation Pathways Controlling Reversible Integrin αIIbβ3 Activation

doi: 10.1055/s-0044-1786987

Figure Lengend Snippet: GPVI- and PAR1-induced changes in platelet morphology. Washed platelets were treated with RO-318425 (10 µM), GF109203X (3 µM), or ticagrelor (1 µM) for 10 minutes before ( A ) or after ( B ) stimulation with 5 µg/mL CRP-XL or 15 µM TRAP6 (or vehicle as a control) for 10 minutes. Platelet morphology was imaged using an inverted light transmission microscope with 63x objective, scale bar = 10 µm, all images are the same size. ( C ) Morphology was assessed for percentage of platelets with disc-shape (‘discoid’), intermediate phenotype (“intermediate”), or formed filopodia and lamellipodia (“filopodia”). Means ± SD ( n = 3–4); Chi-square test, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 for inhibitor vs. control condition (none) without inhibitor at endpoint.

Article Snippet: The platelet agonist thrombin receptor-activating peptide 6, TRAP6 (SFLLRN, PAR1), was purchased from Bachem (Bubendorf, Switzerland); cross-linked CRP was obtained from CambCol (Cambridge, United Kingdom); the PAR4 agonist AYPGKF was from Bio-Techne (Minneapolis, Minnesota, United States).

Techniques: Control, Transmission Assay, Microscopy

Journal: bioRxiv

Article Title: Autocrine Proteinase Activated Receptor (PAR) mediated signaling in prostate cancer cells

doi: 10.1101/2022.08.22.504840

Figure Lengend Snippet: Representative traces of calcium signalling in WT, PAR1-KO, and PAR2-KO PC3 cells treated with PAR1 agonists ( A ) thrombin (0.03 – 10 U ml - ) and ( B ) TFLLR-NH2 (1 – 300 µM), PAR2 agonists ( C ) trypsin (0.3 – 100 nM) and ( D ) SLIGRL-NH2 (1 – 300 µM), and PAR4 agonist ( E ) AYPGKF-NH2 (1 – 300 µM).

Article Snippet: PARs activating peptides TFLLR-NH 2 (PAR1), SLIGRL-NH 2 (PAR2) and AYPGKF-NH 2 (PAR4) (≥95% purity by HPLC) were synthesized by GenScript.

Techniques: