Journal: Future Science OA

Article Title: PAR2 regulates proliferation, migration of lung cancer and chemotherapy sensitivity by involving PTEN pathway

doi: 10.1080/20565623.2025.2535221

Figure Lengend Snippet: PAR2 expression level in lung cancer cells transfected either with pcDNA3-PAR2 or PAR2 shRNA. (A) PAR2 expression level after A549 cells were transfected with pcDNA3-PAR2; (B) PAR2 expression level after H1299 cells were transfected with pcDNA3-PAR2; (C) PAR2 expression level after A549 cells were transfected with PAR2 shRNA; (D) PAR2 expression level after H1299 cells were transfected with PAR2 shRNA.

Article Snippet: Human PAR2 sandwich enzyme-linked immunosorbent assay (ELISA) kit (Novus Biologicals, USA) was used to detect serum PAR2 level according to the manufacture’s instructions.

Techniques: Expressing, Transfection, shRNA

Journal: Future Science OA

Article Title: PAR2 regulates proliferation, migration of lung cancer and chemotherapy sensitivity by involving PTEN pathway

doi: 10.1080/20565623.2025.2535221

Figure Lengend Snippet: Overexpression PAR2 promoted growth of lung cancer cells with or without paclitaxel. (A–D) The growth of A549 cells with or without paclitaxel after transfection with pcDNA3-PAR2; (E–H) The growth of H1299 cells with or without paclitaxel after transfection with pcDNA3-PAR2.

Article Snippet: Human PAR2 sandwich enzyme-linked immunosorbent assay (ELISA) kit (Novus Biologicals, USA) was used to detect serum PAR2 level according to the manufacture’s instructions.

Techniques: Over Expression, Transfection

Journal: Future Science OA

Article Title: PAR2 regulates proliferation, migration of lung cancer and chemotherapy sensitivity by involving PTEN pathway

doi: 10.1080/20565623.2025.2535221

Figure Lengend Snippet: Knockdown PAR2 decreased growth of lung cancer cells with or without paclitaxel. (A–D) The growth of A549 cells with or without paclitaxel after transfection with PAR2 shRNA; (E–H) The growth of H1299 cells with or without paclitaxel after transfection with PAR2 shRNA.

Article Snippet: Human PAR2 sandwich enzyme-linked immunosorbent assay (ELISA) kit (Novus Biologicals, USA) was used to detect serum PAR2 level according to the manufacture’s instructions.

Techniques: Knockdown, Transfection, shRNA

Journal: Future Science OA

Article Title: PAR2 regulates proliferation, migration of lung cancer and chemotherapy sensitivity by involving PTEN pathway

doi: 10.1080/20565623.2025.2535221

Figure Lengend Snippet: PAR2 inhibited paclitaxel-associated apoptosis in lung cancer cells. (A) Caspase 3/7 activity in A549 cell transfected with pcDNA3-PAR2; (B) Caspase 3/7 activity in H1299 cell transfected with pcDNA3-PAR2; (C–F) Bcl-2 and BAX expression in A549 and H1299 cells.

Article Snippet: Human PAR2 sandwich enzyme-linked immunosorbent assay (ELISA) kit (Novus Biologicals, USA) was used to detect serum PAR2 level according to the manufacture’s instructions.

Techniques: Activity Assay, Transfection, Expressing

Journal: Future Science OA

Article Title: PAR2 regulates proliferation, migration of lung cancer and chemotherapy sensitivity by involving PTEN pathway

doi: 10.1080/20565623.2025.2535221

Figure Lengend Snippet: PAR2 played essential roles in invasion and migration of lung cancer cells. (A, B) up-regulation of PAR2 increased migration and invasion of A549 cells. (C, D) down-regulation of PAR2 decreaased migration and invasion of A549 cells. (E, F) up-regulation of PAR2 increased migration and invasion of H1299 cells. (G, H) down-regulation of PAR2 decreased migration and invasion of H1299 cells.

Article Snippet: Human PAR2 sandwich enzyme-linked immunosorbent assay (ELISA) kit (Novus Biologicals, USA) was used to detect serum PAR2 level according to the manufacture’s instructions.

Techniques: Migration

Journal: Future Science OA

Article Title: PAR2 regulates proliferation, migration of lung cancer and chemotherapy sensitivity by involving PTEN pathway

doi: 10.1080/20565623.2025.2535221

Figure Lengend Snippet: PAR2 altered PTEN/AKT protein expression in lung cancer cells. (A–C) Impact of up-regulation of PAR2 on expression of p-AKT, AKT and PTEN in A549 and H1299 cells. (D–F) Impact of down-regulation of PAR2 on expression of p-AKT, AKT and PTEN in A549 and H1299 cells.

Article Snippet: Human PAR2 sandwich enzyme-linked immunosorbent assay (ELISA) kit (Novus Biologicals, USA) was used to detect serum PAR2 level according to the manufacture’s instructions.

Techniques: Expressing

Journal: Future Science OA

Article Title: PAR2 regulates proliferation, migration of lung cancer and chemotherapy sensitivity by involving PTEN pathway

doi: 10.1080/20565623.2025.2535221

Figure Lengend Snippet: PAR2 expression in human lung cancer tissue. (A) PAR2 levels in different stage of lung cancer tissue and normal lung tissue. (B) Immunohistochemical studies for Ki-67 and PTEN on different levels of PAR2 in lung tissue.

Article Snippet: Human PAR2 sandwich enzyme-linked immunosorbent assay (ELISA) kit (Novus Biologicals, USA) was used to detect serum PAR2 level according to the manufacture’s instructions.

Techniques: Expressing, Immunohistochemical staining

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: Mouse PAR2 (mPAR2) cDNA plasmid was purchased from R&D Systems (catalog number RDC0167) and cloned between the BamHI and XbaI sites of pCDNA3.1/Zeo(+) vector.

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

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. 8 Effect of I-287 on intracellular signaling pathways induced by the two human PAR2 agonists, Trypsin, and SLIGKV-NH2. The pathways inhibited by I-287 are in black, whereas the unaffected pathways are in gray.

Article Snippet: Mouse PAR2 (mPAR2) cDNA plasmid was purchased from R&D Systems (catalog number RDC0167) and cloned between the BamHI and XbaI sites of pCDNA3.1/Zeo(+) vector.

Techniques: Protein-Protein interactions

Journal: The Journal of Biological Chemistry

Article Title: Polarization of protease-activated receptor 2 (PAR-2) signaling is altered during airway epithelial remodeling and deciliation

doi: 10.1074/jbc.RA120.012710

Figure Lengend Snippet: A. fumigatus CM directly activates PAR-2 in airway cells. a, diagram of GPCR interaction with β-arrestin after activation and phosphorylation by G-receptor kinase (GRK) b, diagram of the Trio assay (modified from Ref. 26) used to detect PAR-2 activation. A heterologously-expressed GPCR (in this case PAR-2) is tagged at the C-terminal end with the β11-strand of GFP. The receptor is co-expressed with β-arrestin tagged with the β10-strand of GFP along with soluble GFP β-strands 1–9. Association of the PAR-2 with β-arrestin allows the formation of a complete fluorescent GFP molecule; soluble mCherry is included on the plasmid as a transfection control (not shown in the model). c, representative images of A549 cells expressing PAR-2 Trio components (plus mCherry as transfection control) stimulated with 10 μm 2FLI for 0–120 min. Scale bars are 15 μm. d, normalized GFP fluorescence in cells stimulated with 2FLI or buffer alone (HBSS) at time points taken over 2 h. Data points are independent experiments imaged on three different days (n = 5–9). a and b were created with Biorender.com.

Article Snippet: Cells were co-transfected with two pcDNA3 plasmids encoding the GFP β-strands 1–9 (Addgene catalog no. 121684) and a pcDNA3 plasmid expressing the receptor fused to GFP β-strand 11, an arrestin fused to GFP β-strand 10, and mCherry as a transfection marker (Addgene catalog no. 113609 or 113610) and imaged 24 h after transfection at room temperature using standard GFP and TRITC filters on an Olympus IX83 microscope with ×40 (0.75NA PlanFluor) objective.

Techniques: Activation Assay, Phospho-proteomics, Modification, Plasmid Preparation, Transfection, Control, Expressing, Fluorescence

Journal: Cells

Article Title: KLK6/PAR1 Axis Promotes Tumor Growth and Metastasis by Regulating Cross-Talk between Tumor Cells and Macrophages.

doi: 10.3390/cells11244101

Figure Lengend Snippet: Figure 6. KLK6 stimulated TNF-α production in macrophages via PAR1. (A–D) RAW 264.7 cells were transfected with control siRNA, two PAR1 siRNAs, or two PAR2 siRNAs. (A) PAR1 and PAR2 mRNA levels in the indicated siRNAs-transfected cells were measured by RT-PCR. (B) The level of secreted TNF-α in the supernatants of the indicated siRNAs-transfected cells was analyzed by ELISA. (C,D) B16F10 and LLC cells were treated with the CMs of the indicated siRNA-transfected RAW 264.7 cells for 24 h. The mRNA (C) and secreted protein (D) of CXCL1 in B16F10 and LLC cells treated with the CMs of indicated RAW 264.7 cells were analyzed by RT-PCR and ELISA, respectively. (E–G) RAW 264.7 cells were treated with DMSO (as control), a PAR1 antagonist, or a PAR2 antagonist for 24 h. (E) The level of secreted TNF-α in the supernatants of the indicated RAW 264.7 cells was analyzed by ELISA. (F) The level of CXCL1 mRNA in the B16F10 and LLC cells treated with the CMs of indicated RAW 264.7 cells was analyzed by RT-PCR. (G) The level of secreted CXCL1 in the supernatants of B16F10 cells treated with the CMs of the indicated RAW 264.7 cells was measured by ELISA. Statistical significance was determined by the Student’s t-test. * p < 0.05 and ** p < 0.01. Data are representative of three experiments.

Article Snippet: CXCL1 neutralizing antibody (R&D Systems, Minneapolis, MN, USA, MAB453), CXCL1 (R&D Systems, AF-453), PAR1 antagonist (Med-ChemExpress, Monmouth Junction, NJ, USA, SCH 79797), and PAR2 antagonist (Axon-Medchem, Reston, VA, USA, Axon 1622) were purchased from R&D Systems, MedChemExpress and AxonMedchem, respectively. β-actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA, sc47778), Ki67 (Santa Cruz, sc-550609), GFP (Santa Cruz, sc-9996), PAR1 (Novus Biologicals, Novus, Centennial, CO, USA, NBP1-71770), and PAR2 (Abcam, Cambridge, UK, ab180953) antibodies were obtained from Santa Cruz Biotechnology, Novus Biologicals and Abcam, respectively. β-actin was used to normalize the level of proteins.

Techniques: Transfection, Control, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay