Journal: Data in Brief

Article Title: Data regarding the effects of thrombin and dabigatran-inhibited thrombin on protease-activated receptor 1 and activation of human atrial fibroblasts

doi: 10.1016/j.dib.2018.05.124

Figure Lengend Snippet: (A) Representative pictures of primary human atrial Fib from patients without AF, immunostained for PAR1 after no treatment or exposure to thrombin (Th), dabigatran followed by thrombin (Dab + Th), or dabigatran alone (Dab). ( B ) Representative pictures of immunofluorescence for PAR1 in primary Fib from non-fibrillating atria, transfected with negative control siRNA (siCTR) or siRNA targeting human PAR1 (siPAR1), with or without subsequent incubation with dabigatran (Dab). Magnification is 200×, bars indicate 50 μm. ( C ) Western blot for PAR1, confirming the knockdown of PAR1 protein after transfection with siPAR1, but not siCTR.

Article Snippet: Cells were incubated for 1 h with a mouse monoclonal primary antibody against Arg27-Thr102 and Ser375-Thr425 of human PAR1 (clone 731115, R&D Systems, Minneapolis, MN, USA), then for 30 min with an Alexa Fluor® 488-conjugated secondary antibody (ThermoFisher Scientific).

Techniques: Immunofluorescence, Transfection, Negative Control, Incubation, Western Blot, Knockdown

Journal: Data in Brief

Article Title: Data regarding the effects of thrombin and dabigatran-inhibited thrombin on protease-activated receptor 1 and activation of human atrial fibroblasts

doi: 10.1016/j.dib.2018.05.124

Figure Lengend Snippet: Representative images ( A ) and quantification ( B ) of immunofluorescence for αSMA in primary human atrial Fib left untreated (CTR), incubated with thrombin (Th) and/or dabigatran (Dab), or transfected with a control (siCTR) or a PAR1-targeting (siPAR1) siRNA with or without subsequent exposure to Th or Dab as indicated. Fib were obtained from subjects with no history of AF. Magnification 200×, bars 50 μm. Data in B are from three independent replicates and were compared by the Kruskal-Wallis test.

Article Snippet: Cells were incubated for 1 h with a mouse monoclonal primary antibody against Arg27-Thr102 and Ser375-Thr425 of human PAR1 (clone 731115, R&D Systems, Minneapolis, MN, USA), then for 30 min with an Alexa Fluor® 488-conjugated secondary antibody (ThermoFisher Scientific).

Techniques: Immunofluorescence, Incubation, Transfection, Control

Journal: Data in Brief

Article Title: Data regarding the effects of thrombin and dabigatran-inhibited thrombin on protease-activated receptor 1 and activation of human atrial fibroblasts

doi: 10.1016/j.dib.2018.05.124

Figure Lengend Snippet: Primary atrial Fib from subjects without AF were left untreated (CTR), exposed to thrombin (Th) and/or dabigatran (Dab), or transfected with a control (siCTR) or a PAR1-targeting (siPAR1) siRNA with or without subsequent incubation with Th or Dab. At the end of treatments, cells were stained with Picro-Sirius red. Representative images of the staining in conventional and polarized light microscopy are given in ( A ) and ( B ), respectively, while quantification of the red signal in conventional light microscopy is presented in ( C ). Magnification is 200× and bars correspond to 50 μm. Data in the graph are from three independent replicates and were compared by using the Kruskal-Wallis test.

Article Snippet: Cells were incubated for 1 h with a mouse monoclonal primary antibody against Arg27-Thr102 and Ser375-Thr425 of human PAR1 (clone 731115, R&D Systems, Minneapolis, MN, USA), then for 30 min with an Alexa Fluor® 488-conjugated secondary antibody (ThermoFisher Scientific).

Techniques: Transfection, Control, Incubation, Staining, Light Microscopy

Journal: Data in Brief

Article Title: Data regarding the effects of thrombin and dabigatran-inhibited thrombin on protease-activated receptor 1 and activation of human atrial fibroblasts

doi: 10.1016/j.dib.2018.05.124

Figure Lengend Snippet:

Article Snippet: Cells were incubated for 1 h with a mouse monoclonal primary antibody against Arg27-Thr102 and Ser375-Thr425 of human PAR1 (clone 731115, R&D Systems, Minneapolis, MN, USA), then for 30 min with an Alexa Fluor® 488-conjugated secondary antibody (ThermoFisher Scientific).

Techniques: Microscopy, Fluorescence, Light Microscopy, Software, Knockdown, Expressing, Immunofluorescence, Western Blot, Staining, Activation Assay

Journal: American Journal of Respiratory and Critical Care Medicine

Article Title: Pulmonary Epithelium Is a Prominent Source of Proteinase-activated Receptor-1–inducible CCL2 in Pulmonary Fibrosis

doi: 10.1164/rccm.200712-1827oc

Figure Lengend Snippet: Figure 1. Proteinase-activated receptor-1 (PAR1) and che- mokine (CC motif) ligand-2 (CCL2) are coexpressed in bleomycin-induced lung injury. Shown is the immunohis- tochemical localization of PAR1 (A, C, and E) and CCL2 (B, D, and F) in adjacent lung sections 6 days after instillation of saline (A and B) or bleomycin (C–F). Increased immu- noreactivity for both PAR1 and CCL2 was observed on bronchial epithelial cells (BE, arrows) and alveolar epithe- lial cells (AE) as well as macrophages (M) in both moderate (C and D) and severe (E and F) lesions. Scale bars: (A–F) 50 mm.

Article Snippet: Immunostaining was undertaken by the avidin– biotinylated enzyme complex method (Vector Laboratories, Burlingame, CA) with antibodies against human PAR1 (anti-TRED; 3 mg/ml), mouse PAR1 (anti-SFFL; 0.14 mg/ml), human CCL2 (MAB2791; final concentration, 8 mg/ml; R&D Systems), or mouse CCL2 (sc-1784; final concentration, 0.5 mg/ml; Santa Cruz Biotechnology), and equivalent concentrations of polyclonal nonimmune IgG controls.

Techniques: Saline

Journal: American Journal of Respiratory and Critical Care Medicine

Article Title: Pulmonary Epithelium Is a Prominent Source of Proteinase-activated Receptor-1–inducible CCL2 in Pulmonary Fibrosis

doi: 10.1164/rccm.200712-1827oc

Figure Lengend Snippet: Figure 2. Proteinase-activated receptor-1 (PAR1) and chemokine (CC motif) ligand-2 (CCL2) immunostain- ing in idiopathic pulmonary fibrosis (IPF) identifies epithelial cells and macrophages in fibrotic areas as major cell types displaying dual immunoreactivity. Shown is immunohistochemistry for the epithelial cell marker cytokeratin-7 (CK7) (A and B), PAR1 (C and D), CCL2 (E and F), and CD68 (G and H), in serial sections of IPF (right) and control lung (left). Immunostaining for CCL2 (F), observed in discrete areas of the IPF lung, is associated with both the epithelial marker CK7 (B) and PAR1 (D), most prominently on cells morphologically characteristic of hyperplastic alveolar (asterisks) and bronchial epithelium (arrowheads) (B, D, and F). PAR1 and CCL2 staining is also evident in regions of macro- phage infiltration demarcated by CD68 immunoreactiv- ity (D, F, and H). Control human lung is immunoreactive for PAR1 (C) in CK7-positive alveolar and bronchial epithelial cells (A). CCL2 staining is absent from all CK7- positive cells in control lung (A and E); however, weak expression is observed in CD68-positive cells morpho- logically characteristic of macrophages (G), which are positive for PAR1 (C). Scale bars: (A–H) 100 mm.

Article Snippet: Immunostaining was undertaken by the avidin– biotinylated enzyme complex method (Vector Laboratories, Burlingame, CA) with antibodies against human PAR1 (anti-TRED; 3 mg/ml), mouse PAR1 (anti-SFFL; 0.14 mg/ml), human CCL2 (MAB2791; final concentration, 8 mg/ml; R&D Systems), or mouse CCL2 (sc-1784; final concentration, 0.5 mg/ml; Santa Cruz Biotechnology), and equivalent concentrations of polyclonal nonimmune IgG controls.

Techniques: Immunohistochemistry, Marker, Control, Immunostaining, Staining, Expressing

Journal: American Journal of Respiratory and Critical Care Medicine

Article Title: Pulmonary Epithelium Is a Prominent Source of Proteinase-activated Receptor-1–inducible CCL2 in Pulmonary Fibrosis

doi: 10.1164/rccm.200712-1827oc

Figure Lengend Snippet: Figure 3. Dual immunofluorescence reveals coimmunos- taining for proteinase-activated receptor-1 (PAR1) and chemokine (CC motif) ligand-2 (CCL2) on epithelial cells overlying fibrotic foci and infiltrating macrophages. Im- munohistochemistry for the epithelial cell marker cytoker- atin-7 (CK7) (A) and the macrophage marker CD68 (E) in serial sections, directly adjacent to dual immunofluores- cence costaining for PAR1 and CCL2 (C). PAR1-positive cells (B) are visualized in red, CCL2 (D) in green, and cells copositive for PAR1 and CCL2 appear orange (C). Cell nuclei were visualized by 49,6-diamidino-2-phenylindole (DAPI) staining (F, blue). Colocalization of PAR1 and CCL2 is prominent on CK7-positive epithelial cells (B and A) overlying the fibrotic focus, in addition to infiltrating inflammatory cells positive for the macrophage antigen CD68 (B and E). Scale bars: (A and E) 50 mm.

Article Snippet: Immunostaining was undertaken by the avidin– biotinylated enzyme complex method (Vector Laboratories, Burlingame, CA) with antibodies against human PAR1 (anti-TRED; 3 mg/ml), mouse PAR1 (anti-SFFL; 0.14 mg/ml), human CCL2 (MAB2791; final concentration, 8 mg/ml; R&D Systems), or mouse CCL2 (sc-1784; final concentration, 0.5 mg/ml; Santa Cruz Biotechnology), and equivalent concentrations of polyclonal nonimmune IgG controls.

Techniques: Marker, Staining

Journal: American Journal of Respiratory and Critical Care Medicine

Article Title: Pulmonary Epithelium Is a Prominent Source of Proteinase-activated Receptor-1–inducible CCL2 in Pulmonary Fibrosis

doi: 10.1164/rccm.200712-1827oc

Figure Lengend Snippet: Figure 4. Proteinase-activated receptor-1 (PAR1) and chemokine (CC motif) ligand-2 (CCL2) mRNA levels are increased in alveolar septae in idiopathic pulmonary fibrosis (IPF). (A) PAR1 and monocyte chemotac- tic protein (MCP)-1 expression in microdissected alveolar septae from human IPF and donor lungs as analyzed by quantitative real-time polymerase chain reaction. Data are enumerated as the fold increase relative to mean mRNA levels in donors, normalized to the housekeep- ing gene hypoxanthine phosphoribosyltransferase (HPRT) 6 SEM (patients with IPF: MCP-1, n 5 8; PAR1, n 5 9) (donor subjects: MCP-1, n 5 7; PAR1, n 5 6). *P , 0.05, **P , 0.005 (Student t test, two-tailed, two sample, equal variance). B and C show a typical example of an alveolar septum (B) before and (C) after microdissection.

Article Snippet: Immunostaining was undertaken by the avidin– biotinylated enzyme complex method (Vector Laboratories, Burlingame, CA) with antibodies against human PAR1 (anti-TRED; 3 mg/ml), mouse PAR1 (anti-SFFL; 0.14 mg/ml), human CCL2 (MAB2791; final concentration, 8 mg/ml; R&D Systems), or mouse CCL2 (sc-1784; final concentration, 0.5 mg/ml; Santa Cruz Biotechnology), and equivalent concentrations of polyclonal nonimmune IgG controls.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Two Tailed Test, Laser Capture Microdissection

Journal: iScience

Article Title: Coagulation factors promote brown adipose tissue dysfunction and abnormal systemic metabolism in obesity

doi: 10.1016/j.isci.2022.104547

Figure Lengend Snippet:

Article Snippet: anti-PAR1 antibody , Novus Biologicals , NBP1-71770 RRID: AB_11027203.

Techniques: Virus, Expressing, Clinical Proteomics, Recombinant, Coagulation, Enzyme-linked Immunosorbent Assay, Microarray, Software