Journal: Oncology Reports

Article Title: The expression and the functional roles of tissue factor and protease-activated receptor-2 on SW620 cells

doi: 10.3892/or_00000111

Figure Lengend Snippet: Figure 2. PAR1 and PAR2 antigen level on SW620 and SW480 cells detected by immunocytochemistry using monoclonal antibodies. A and C, SW480 cells; B and D, SW620 cells; A and B, reaction with anti-PAR1 antibody (α-R1); C and D, reaction with anti-PAR2 antibody (α-R2). The pictures were taken under light microscopy with high power field (x400).

Article Snippet: PAR1 agonist SFLLRN-NH2, PAR2 agonist SLIGKV-NH2 and antagonist VKGILS-NH2 were synthesized by Proteintech Group Inc. (Wuhan, P.R.

Techniques: Immunocytochemistry, Bioprocessing, Light Microscopy

Journal: Oncology Reports

Article Title: The expression and the functional roles of tissue factor and protease-activated receptor-2 on SW620 cells

doi: 10.3892/or_00000111

Figure Lengend Snippet: Figure 3. PAR1 and PAR2 mRNA expression on SW620 and SW480 cells investigated by RT-PCR. Total cellular RNA was extracted from the cells (1x106) and RT-PCR was carried out using PAR1, PAR2 primes respectively. The RT-PCR products were visualized on 2% agarose gel stained with ethidium bromide and photographed under UV light.

Article Snippet: PAR1 agonist SFLLRN-NH2, PAR2 agonist SLIGKV-NH2 and antagonist VKGILS-NH2 were synthesized by Proteintech Group Inc. (Wuhan, P.R.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Staining

Journal: Oncology Reports

Article Title: The expression and the functional roles of tissue factor and protease-activated receptor-2 on SW620 cells

doi: 10.3892/or_00000111

Figure Lengend Snippet: Figure 4. SW620 cell migratory potential measured by modified Boyden chambers. The cells were treated with different agents and the migration assay was carried out as described in Materials and methods. A, The effects of TF/VIIa; B, The effects of PAR1 and PAR2; C, The correlation of TF/VIIa and PAR2. Cells without any agent were used as a control; VIIa, activated factor VII (10 nM); α-TF, anti-TF antibody; α-R1 and α-R2, anti- PAR1 and anti-PAR2 antibodies; mopc-21, isotype control antibody (all the antibodies are monoclonal mouse anti-human, 10 μg/ml); R2-anta, PAR2 antagonist (100 μM); R1-AP and R2-AP, PAR1 and PAR2 agonists (100 μM). Data shown are the mean ± SEM of ten fields of triplicate determinations.

Article Snippet: PAR1 agonist SFLLRN-NH2, PAR2 agonist SLIGKV-NH2 and antagonist VKGILS-NH2 were synthesized by Proteintech Group Inc. (Wuhan, P.R.

Techniques: Modification, Migration, Control

Journal: Oncology Reports

Article Title: The expression and the functional roles of tissue factor and protease-activated receptor-2 on SW620 cells

doi: 10.3892/or_00000111

Figure Lengend Snippet: Figure 5. SW620 cell proliferation potential detected using [3H]-TdR incorporation. The cells were treated with different agents and the proliferation potential was investigated by [3H]-TdR incorporation. A, The effects of TF/VIIa; B, The effects of PAR1 and PAR2; C, The correlation of TF/VIIa and PAR2. Cells without any agent were used as a control. All the concentration of materials was similar to that in Fig. 4. Data shown are the mean ± SEM of triplicate determinations. *P<0.01 vs. control; **P<0.001 vs control; LP <0.01 vs. R2-AP alone and #P<0.01 vs. VIIa alone.

Article Snippet: PAR1 agonist SFLLRN-NH2, PAR2 agonist SLIGKV-NH2 and antagonist VKGILS-NH2 were synthesized by Proteintech Group Inc. (Wuhan, P.R.

Techniques: Control, Concentration Assay

Journal: Oncology Reports

Article Title: The expression and the functional roles of tissue factor and protease-activated receptor-2 on SW620 cells

doi: 10.3892/or_00000111

Figure Lengend Snippet: Figure 6. IL-8 secretion by SW620 cells treated with various materials. The cells were treated with materials similar to that of migratory assay and IL-8 protein level in conditioned media was detected using the IL-8 ELISA kit. A, The effects of TF/VIIa; B, The effects of PAR1 and PAR2; C, The correlation of TF/VIIa and PAR2. Cells without any agent were used as a control. Data shown are the mean ± SEM of triplicate determinations.

Article Snippet: PAR1 agonist SFLLRN-NH2, PAR2 agonist SLIGKV-NH2 and antagonist VKGILS-NH2 were synthesized by Proteintech Group Inc. (Wuhan, P.R.

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Communications biology

Article Title: The PAR2 inhibitor I-287 selectively targets Gα q and Gα 12/13 signaling and has anti-inflammatory effects.

doi: 10.1038/s42003-020-01453-8

Figure Lengend Snippet: Fig. 5 I-287 inhibits PAR2-mediated activation of DAG/Ca2+/PKC and RhoA/SRF-RE, as well as FAK and ERK1/2 signaling pathways. a, b Impact of increasing concentrations of I-287 (15 min) on DAG production (a) and PKC activation (b) induced after 1 (DAG) or 5 (PKC) min stimulation with an EC80 concentration of hTrypsin or SLIGKV-NH2 in HEK293 cells co-expressing hPAR2 and the indicated unimolecular BRET2-based biosensors. Results are expressed as ΔBRET in % of the response induced by EC80 of respective agonists in the absence of I-287 (mean ± SEM; n = 4–5). c Impact of increasing concentrations of I-287 (30 min) on intracellular Ca2+ mobilization induced by an EC80 concentration of hTrypsin or SLIGKV-NH2 in HEK293 cells endogenously expressing hPAR2. Results are expressed as % of the response induced by respective agonists in the absence of I-287 (mean ± SEM; n = 3-4). d Impact of I-287 (10 µM, 30 min) on hPAR2-promoted SRF-RE reporter gene activation induced after 6 h stimulation with hTrypsin (10 U/mL) or SLIGKV-NH2 (100 µM) in HEK293 cells expressing hPAR2. FBS (10%) was used as control. Results are expressed as % of the response induced by respective agonists in the absence of I-287 (mean ± SEM; n = 3–5; unpaired t-test: *p < 0.05 and **p < 0.01 compared to respective control cells, ns: nonsignificant). e, f Kinetics of FAK and ERK1/2 phosphorylation in HEK293 cells expressing hPAR2 and pretreated with DMSO or I-287 (10 µM, 30 min) before stimulation with hTrypsin (1 U/mL) or SLIGKV-NH2 (100 µM) at the indicated times. Representative immunoblots of FAK and ERK1/2 phosphorylation are shown. Western blots were quantified and expressed as the ratio of phosphorylated protein level (P-FAK or P-ERK1/2) normalized over total protein (t-FAK or t-ERK1/2; mean ± SEM; n = 3–5; two-way ANOVA followed by Tukey’s post hoc test: *p < 0.05, **p < 0.01, and ***p < 0.001 compared to DMSO-treated cells at the respective time).

Article Snippet: Plasmids. hPAR2 cDNA plasmid was purchased from Origen (IBD90.1 clone; catalog number SC322345), where the serine in position 291 has been mutated in threonine (S291T) to reproduce the hPAR2 phenotype observed in HEK293 and HCT 116 cells.

Techniques: Activation Assay, Protein-Protein interactions, Concentration Assay, Expressing, Control, Phospho-proteomics, Western Blot

Journal: British Journal of Pharmacology

Article Title: Newly developed serine protease inhibitors decrease visceral hypersensitivity in a post‐inflammatory rat model for irritable bowel syndrome

doi: 10.1111/bph.14396

Figure Lengend Snippet: TaqMan primers used for qPCR analysis of colonic and DRG samples

Article Snippet: Table A1 Protein Gene ID Tryptase αβ1 Rn00570928_m1 Matriptase Rn00586242_m1 Cathepsin G Rn01489144_g1 Urokinase plasminogen activator Rn00565261_m1 KLK2 Rn00820615_m1 KLK4 Rn01498534_g1 KLK8 Rn01476995_m1 PAR2 Rn00588089_m1 PAR4 Rn00587480_m1 TRPA1 Rn01473803_m1 TRPV1 Rn00583117_m1 TRPV4 Rn00576745_m1 GAPDH Rn01775763_g1 β‐actin Rn00667869_m1 Open in a separate window TaqMan primers used for qPCR analysis of colonic and DRG samples Table A2 Proteases UAMC‐00050 UAMC‐01162 Nafamostat mesilate IC 50 (μM) IC 50 (μM) Est.

Techniques: