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: Frontiers in Molecular Biosciences

Article Title: The predictive role of protease-activated receptor (PAR-1) polymorphisms and activated microplatelets on the severity of atherosclerosis – preliminary studies

doi: 10.3389/fmolb.2025.1662954

Figure Lengend Snippet: The cytometric analysis (SSC and FCS) of isolated human resting platelets (A, D) , gated and labeled PAR-1 without activation (B, E) , and gated and labeled PAR-1 with activation by 10 µM TRAP (C, F) ; labeled with anti-CD61-FITC and PAR-1-APC antibodies (G–I) . The level of PAR-1 expression was read from gates P1. Markers M1 and M2 indicate the gates for microparticles and normal platelets, with the PAR-1 analysis applied to the summed population. An example image from a patient with DM is shown.

Article Snippet: Next: For the PAR-1 test without PLT activation, 5 μL of PAR-1-APC antibodies at a concentration of 5 μg/5 × 10 5 cells (Allophycocyanin (APC)-conjugated mouse monoclonal anti-human PAR-1; clone# 731115; mouse isotype: IgG1, R&D Systems, Minneapolis, Canada) and 5 μL of CD61-FITC antibodies (Monoclonal Mouse Anti-Human CD61, Platelet Glycoprotein IIIa/FITC, Clone Y2/51, code: F0803, DakoCytomation, Glostrup, Denmark) were added.

Techniques: Isolation, Labeling, Activation Assay, Expressing

Journal: Frontiers in Molecular Biosciences

Article Title: The predictive role of protease-activated receptor (PAR-1) polymorphisms and activated microplatelets on the severity of atherosclerosis – preliminary studies

doi: 10.3389/fmolb.2025.1662954

Figure Lengend Snippet: The percentage of PAR-1 receptor expression before and after the addition of the thrombin receptor activating peptide (TRAP) in blood samples from patients with diabetic macroangiopathy (DM), the control group (CONTROL), and atherosclerosis obliterans (AO).

Article Snippet: Next: For the PAR-1 test without PLT activation, 5 μL of PAR-1-APC antibodies at a concentration of 5 μg/5 × 10 5 cells (Allophycocyanin (APC)-conjugated mouse monoclonal anti-human PAR-1; clone# 731115; mouse isotype: IgG1, R&D Systems, Minneapolis, Canada) and 5 μL of CD61-FITC antibodies (Monoclonal Mouse Anti-Human CD61, Platelet Glycoprotein IIIa/FITC, Clone Y2/51, code: F0803, DakoCytomation, Glostrup, Denmark) were added.

Techniques: Expressing, Control

Journal: Frontiers in Molecular Biosciences

Article Title: The predictive role of protease-activated receptor (PAR-1) polymorphisms and activated microplatelets on the severity of atherosclerosis – preliminary studies

doi: 10.3389/fmolb.2025.1662954

Figure Lengend Snippet: (A) Separation of DNA molecules in a 3% agarose gel of PAR-1 gene amplification products with the −506 I/D polymorphism. Lanes: 1 – homozygous I/I, 2 – heterozygous I/D, 3 – homozygous D/D, M–GeneRuler™ 50bp DNA Ladder (Fermentas). (B) The percentage distribution of the −506 I/D polymorphism variants in the PAR-1 gene: homozygous D/D (blue), heterozygous I/D (red), and homozygous I/I (green).

Article Snippet: Next: For the PAR-1 test without PLT activation, 5 μL of PAR-1-APC antibodies at a concentration of 5 μg/5 × 10 5 cells (Allophycocyanin (APC)-conjugated mouse monoclonal anti-human PAR-1; clone# 731115; mouse isotype: IgG1, R&D Systems, Minneapolis, Canada) and 5 μL of CD61-FITC antibodies (Monoclonal Mouse Anti-Human CD61, Platelet Glycoprotein IIIa/FITC, Clone Y2/51, code: F0803, DakoCytomation, Glostrup, Denmark) were added.

Techniques: Agarose Gel Electrophoresis, Amplification

Journal: Frontiers in Molecular Biosciences

Article Title: The predictive role of protease-activated receptor (PAR-1) polymorphisms and activated microplatelets on the severity of atherosclerosis – preliminary studies

doi: 10.3389/fmolb.2025.1662954

Figure Lengend Snippet: (A) The result of the restriction digestion of PCR products with the MvaI enzyme to check for the presence of the −1426 C/T polymorphism in the PAR-1 gene. Lanes: 1 – 6 homozygotes C/C, M–GeneRuler™ 100bp DNA Ladder (Fermentas). (B) The percentage distribution of the variants of the −1426 C/T polymorphism in the PAR-1 gene: homozygote C/C (blue color), heterozygote C/T (red color), homozygote T/T (green color).

Article Snippet: Next: For the PAR-1 test without PLT activation, 5 μL of PAR-1-APC antibodies at a concentration of 5 μg/5 × 10 5 cells (Allophycocyanin (APC)-conjugated mouse monoclonal anti-human PAR-1; clone# 731115; mouse isotype: IgG1, R&D Systems, Minneapolis, Canada) and 5 μL of CD61-FITC antibodies (Monoclonal Mouse Anti-Human CD61, Platelet Glycoprotein IIIa/FITC, Clone Y2/51, code: F0803, DakoCytomation, Glostrup, Denmark) were added.

Techniques:

Journal: Frontiers in Molecular Biosciences

Article Title: The predictive role of protease-activated receptor (PAR-1) polymorphisms and activated microplatelets on the severity of atherosclerosis – preliminary studies

doi: 10.3389/fmolb.2025.1662954

Figure Lengend Snippet: (A) Example separations of amplification products of the DNA fragment encompassing the IVSn-14 A/T polymorphism site of the PAR-1 gene using the SNaPshot method. Alleles were determined based on the size of primers and the colors of fluorescently labeled ddNTPs (terminators) incorporated during the primer extension reaction. (A) red peak, wild-type homozygote (AA); (B) green and red peaks, heterozygote (AT); (C) green peak, mutated homozygote (TT). (B) The percentage distribution of the variants of the IVS-14 A/T polymorphism of the PAR-1 gene is as follows: homozygote A/A (blue color), heterozygote A/T (red color), and homozygote T/T (green color).

Article Snippet: Next: For the PAR-1 test without PLT activation, 5 μL of PAR-1-APC antibodies at a concentration of 5 μg/5 × 10 5 cells (Allophycocyanin (APC)-conjugated mouse monoclonal anti-human PAR-1; clone# 731115; mouse isotype: IgG1, R&D Systems, Minneapolis, Canada) and 5 μL of CD61-FITC antibodies (Monoclonal Mouse Anti-Human CD61, Platelet Glycoprotein IIIa/FITC, Clone Y2/51, code: F0803, DakoCytomation, Glostrup, Denmark) were added.

Techniques: Amplification, Labeling

Journal: Frontiers in Molecular Biosciences

Article Title: The predictive role of protease-activated receptor (PAR-1) polymorphisms and activated microplatelets on the severity of atherosclerosis – preliminary studies

doi: 10.3389/fmolb.2025.1662954

Figure Lengend Snippet: Multivariate analysis: (A–D) Number of microparticles with PAR-1+TRAP; (E–F) Number of microparticles with BMI (Figure 4.12E-F) and age with smoking.

Article Snippet: Next: For the PAR-1 test without PLT activation, 5 μL of PAR-1-APC antibodies at a concentration of 5 μg/5 × 10 5 cells (Allophycocyanin (APC)-conjugated mouse monoclonal anti-human PAR-1; clone# 731115; mouse isotype: IgG1, R&D Systems, Minneapolis, Canada) and 5 μL of CD61-FITC antibodies (Monoclonal Mouse Anti-Human CD61, Platelet Glycoprotein IIIa/FITC, Clone Y2/51, code: F0803, DakoCytomation, Glostrup, Denmark) were added.

Techniques:

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: Coagulation factors and protease-activated receptors are upregulated in brown adipose tissue under metabolic stress (A) Hematoxylin and eosin (HE) staining of brown adipose tissue (BAT) from wild-type mice fed a normal chow (NC) or a high-fat diet (HFD). In the HFD group, mice were fed the diet from 4 weeks of age and were analyzed at 19 to 22 weeks of age. Scale bar = 100 μm. (B and C) Enzyme-linked immunosorbent assay (ELISA; B, n = 10, 10) or immunofluorescent staining (C) for tissue factor in BAT of the indicated mice aged 19-22weeks. Scale bar = 50 μm. (D-H) Results of quantitative polymerase chain reaction (PCR) for coagulation factor VII ( F7 ; D, n = 8, 7) or coagulation factor X ( F10 ; G, n = 8, 7) in mice aged 19-22weeks. Results of enzyme-linked immunosorbent assay (ELISA; E, H) or immunofluorescent staining (F) for coagulation factor VII (E and F, n = 10, 11) or activated coagulation factor X (Factor Xa; H, n = 9, 9) in BAT of the indicated mice aged 19-22weeks. Scale bar = 50 μm. (I) Western blot analysis of protease-activated receptor-1 (PAR-1) or PAR-2 expression in epididymal white adipose tissue (eWAT) or BAT from the indicated groups. Right panel indicates the quantification of protease-activated receptor-1 (PAR-1) relative to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in BAT (n = 5, 5). (J) Immunofluorescent staining for PAR-1 in BAT from the indicated group. Scale bar = 50 μm. (K) RNA sequence data analyzing F2R in brown adipose tissue (BAT) from individuals with a body mass index (BMI) < 25 (n = 8) or BMI ≥25 (n = 23). Datasets were taken from Jespersen et al. ( https://www.biorxiv.org/content/10.1101/2020.05.07.082057v1 ). (L) Enzyme-linked immunosorbent assay (ELISA) for Factor Xa in plasma from NC or HFD mice aged 19-22weeks (left panel; n = 3, 3), and plasma from lean (BMI <22) or obese (BMI >28) human volunteers (right panel; n = 13, 13). All data were analyzed by a 2-tailed Student’s t test. ∗p < 0.05, ∗∗p < 0.01. Values represent the mean ± SEM NS = not significant. All data are from different biological replicates. See also Figure S1 .

Article Snippet: pCMV6-Entry-mouse F2r , OriGENE , MR206862.

Techniques: Coagulation, Staining, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Sequencing, Clinical Proteomics

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: Brown adipose tissue-specific protease-activated receptor-1 depletion ameliorates brown adipose tissue dysfunction Brown adipose tissue (BAT)-specific protease-activated receptor-1 (PAR1) knockout (KO) mice (UCP1-Cre +/− ; PAR1 flox/flox ; BAT PAR1 KO) were fed a high-fat diet (HFD) from 4 weeks of age and physiological studies were performed after 12 to 13 weeks. Tissues were harvested at 18-21 weeks of age. (A, B, and C) Hematoxylin and eosin (HE) staining (A; scale bar = 100 μm), dihydroethidium (DHE) staining (B; scale bar = 50 μm), and transmission electron microscopy (C; scale bar = 2 μm) of BAT from littermate control mice (PAR1 flox/flox ; Con), BAT PAR1 KO, and BAT PAR1 KO mice treated with an FXa inhibitor (BAT PAR1 KO + FXa-i). Right panels in Figure 4 A indicate the quantification of large lipid droplets (n = 4, 3, 4), and Figure 4 B indicates the quantification of dihydroethidium (DHE) staining (n = 5, 5, 5) of BAT in littermate control mice (PAR1 flox/flox ; Con), BAT PAR1 KO, and BAT PAR1 KO + FXa-i mice. Right panel in Figure 4 C indicates mitochondria area in % (analyzed as mitochondrial area/[non-capillary and non-lipid area]) in the pericapillary area of the respective groups (n = 4, 4, 4). (D, E) Cold tolerance test (CTT; D, n = 16, 8, 5) and glucose tolerance test (GTT; E, n = 14, 11, 11) in the indicated mice aged 13 weeks for CTT, and 12 weeks for GTT. ∗ indicates Con vs. BAT PAR1 KO, ## indicates Con vs. BAT PAR1 KO + FXa-i. Data were analyzed by 2-way ANOVA followed by Tukey’s multiple comparison test (A, B, C) or by 2-way repeated-measures ANOVA followed by Tukey’s multiple comparison test (D, E). ∗p < 0.05, ∗∗p < 0.01, ## p < 0.01. Values represent the mean ± SEM NS = not significant. All data are from different biological replicates. See also Figure S4 .

Article Snippet: pCMV6-Entry-mouse F2r , OriGENE , MR206862.

Techniques: Knock-Out, Staining, Transmission Assay, Electron Microscopy, Control, Comparison

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: Introduction of tissue factor and PAR-1 in brown adipose tissue promotes functional decline of this organ All experiments with Adeno-associated virus (AAV) were performed in mice fed normal chow (NC). AAVs were injected at 10 weeks of age. Physiological studies were performed 10 to 14 days after AAV injection, and tissues were harvested at 14 weeks of age. (A-C) Enzyme-linked immunosorbent assay (ELISA) for tissue factor (A, n = 10, 9) or FactorXa (C, n = 8, 8) in BAT from mice injected with control AAV (Mock) or with both AAV encoding F3 and AAV encoding F2r (AAV F3+F2r) aged 14 weeks. (B) Western blot analysis of PAR-1 in BAT from the indicated groups. Right panel indicates the quantification of PAR1 relative to glyceraldehyde 3-phosphate dehydrogenase (GAPDH; n = 9,9). (D, E, and F) Hematoxylin and eosin (HE) staining (D; scale bar = 100μm), dihydroethidium (DHE) staining (E; scale bar = 50 μm), and transmission electron microscopy (F; scale bar = 10 μm for low magnification [Low Mag] and 2 μm for high magnification [High Mag]) of BAT from the indicated mice. Right panel in Figure 5 E indicates the quantification of dihydroethidium (DHE) analyzed as relative DHE signal (DHE area of AAV F3+F2r/Mock) of indicated mice (n = 4, 5). Right panel in Figure 5 F indicates mitochondria area (%; analyzed as mitochondrial area/[non-capillary and non-lipid area]) in the pericapillary area of the respective groups (n = 4, 4). (G and H) Cold tolerance test (CTT; G, n = 8, 8) and glucose tolerance test (GTT; H, n = 6, 7) in the indicated mice aged 13 weeks for CTT, and 12 weeks for GTT. Data were analyzed by a 2-tailed Student’s t test (A, B, C, E, F) or by 2-way repeated measures ANOVA (G and H). ∗p < 0.05, ∗∗p < 0.01. Values represent the mean ± SEM NS = not significant. All data are from different biological replicates. See also Figure S5 .

Article Snippet: pCMV6-Entry-mouse F2r , OriGENE , MR206862.

Techniques: Functional Assay, Virus, Injection, Enzyme-linked Immunosorbent Assay, Control, Western Blot, Staining, Transmission Assay, Electron Microscopy

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: FactorXa/PAR1 signaling promotes the dysfunction of brown adipocytes (A-H) Nivo Multimode Microplate Reader study analyzing signal of mitochondrial ROS (MitoSox; A, n = 5, 5, 5; C, n = 8, 8, 8; G, n = 8, 8, 8), mitochondrial membrane potential (MitoRed; B, n = 8, 8, 8; D, n = 20, 20, 20; H, n = 8, 8, 8), mitochondrial calcium (Ca 2+ ; Rhod2; E, n = 8, 8, 8; F, n = 8, 8, 8) in the indicated groups. For studies of MitoSox and MitoRed, recombinant FXa protein (10nM) was administered for a total of 3 h and other compounds were administered for 1 h before the administration of FXa at the following concentrations: PAR1 inhibitor (PAR1-i), 1μM; MitoTEMPO, 10μM; MCU inhibitor (250nM). For studies of Rhod2, recombinant FXa was administered for a total of 30 min, with or without the reagents described above. (I) Results of qPCR for expression of coagulation factor VII ( F7 ; n = 5, 6), factor X ( F10 ; n = 6, 6), tissue factor ( F3 ; n = 4, 5), and F2r (PAR-1 transcript; n = 6, 6) in differentiated brown adipocytes incubated with control adenovirus (Mock) or adenovirus encoding constitutively activated Hif-1a (Ad-Hif-1a; 10 MOI, 48 h for F7 , F10, and F2r ; 30 MOI, 24 h for F3 ). (J) Enzyme-linked immunosorbent assay (ELISA) for tissue factor (n = 10, 10), coagulation factor VII (FVII; n = 9, 14), and FXa (n = 11, 9) in conditioned medium from differentiated brown adipocytes incubated with control adenovirus (Mock) or Ad-Hif-1a. (K and L) MitoSox/MitoGreen ratio (K, n = 8, 7, 7) and MitoRed/MitoGreen ratio (L, n = 8, 8, 8) in differentiated brown adipocytes incubated with conditioned medium (CM) obtained from brown adipocytes infected with Mock (Mock-CM), Ad-Hif-1a (Ad-Hif-1a-CM), or Ad-Hif-1a-CM + PAR1-i. Data were analyzed by a 2-tailed Student’s t test (I-L), or by 2-way ANOVA followed by Tukey’s multiple comparison test (A and C-H), or non-parametric Kruskal Wallis test (B). ∗p < 0.05, ∗∗p < 0.01. Values represent the mean ± SEM NS = not significant. All data are from different biological replicates. See also Figure S6 .

Article Snippet: pCMV6-Entry-mouse F2r , OriGENE , MR206862.

Techniques: Membrane, Recombinant, Expressing, Coagulation, Incubation, Control, Enzyme-linked Immunosorbent Assay, Infection, Comparison

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: pCMV6-Entry-mouse F2r , OriGENE , MR206862.

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

Journal: Oncotarget

Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro.

doi: 10.18632/oncotarget.5741

Figure Lengend Snippet: Figure 4: MMP1 induces Ca2+ signalling, and activates migratory proteins and CCIDs in LECs. a. LECs were grown to ~70–80% confluence and then pre-treated with 0.5 μM SCH79797 or solvent (DMSO) and then stimulated with 100 ng/ml activated recombinant MMP1 for 4 h. Cells were lysed, proteins were separated by SDS gel electrophoresis and analysed by Western blotting using the indicated antibodies. Equal sample loading was controlled by Ponceau S staining and ß-actin immunoblotting. Densitometer readings facilitated the comparison of relative protein expression levels with solvent treated control (which was set as “1”). (b) LECs (8 × 103 cells/ well) were pre-treated with 0.5 μM SCH79797 (PAR1 inhibitor) and then charged with FluoForte Dye-loading in presence of SCH79797 for 45 min at 37°C and 15 min at room temperature. Then, cells were stimulated with 100 ng/ml activated recombinant MMP1 for 5 min. Intracellular free calcium was measured with a fluorescence plate reader at 490/525 nm. Experiments were performed in triplicate, error bars indicate means +/− SEM, and asterisks and rhomboids significance (p < 0.05; t-test). (c) Confluent LECs were pre-treated with SCH79797 or solvent (DMSO) for 30 min and then MDA-MB231 spheroids were placed on top of LECs monolayers and co-incubated for 4 h. The areas of CCIDs were analysed using an Axiovert microscope and Zen Little 2012 software.

Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

Techniques: Solvent, Recombinant, SDS-Gel, Electrophoresis, Western Blot, Staining, Comparison, Expressing, Control, Fluorescence, Incubation, Microscopy, Software

Journal: Oncotarget

Article Title: NF-κB contributes to MMP1 expression in breast cancer spheroids causing paracrine PAR1 activation and disintegrations in the lymph endothelial barrier in vitro.

doi: 10.18632/oncotarget.5741

Figure Lengend Snippet: Figure 5: Combined inhibitions of NF-κB and target genes in MDA-MB231 spheroids and LEC monolayers. LECs or MDA-MB231 spheroids were transiently transfected with either non-targeting (n.t.) siRNA or a combination of siRNAs inhibiting the expression of a. NFKB1 & NFKB2, b. or siRNAs inhibiting the expression of ICAM-1 and MMP1 respectively. After 24 h, spheroids were placed on top of confluent LEC monolayers and co-incubated for 4 h. c. MDA-MB231 spheroids were transiently transfected with either non-targeting (n.t.) siRNA or siRNA inhibiting the expression of MMP1. After 24 h, spheroids were placed on top of confluent LEC monolayers, which were pre-treated with 1 μM SCH79797 (inhibitor of PAR1) or solvent (DMSO) for 30 min, and co-incubated for 4 h. The areas of CCIDs were analysed using an Axiovert microscope and Zen Little 2012 software. Experiments were performed in triplicate, error bars indicate means +/− SEM, and asterisks and rhomboids significance (p < 0.05; t-test).

Article Snippet: Polyclonal goat anti-CD54 (ICAM-1) and monoclonal mouse anti-human PAR1 was from R&D system (Minneapolis, MN, USA).

Techniques: Transfection, Expressing, Incubation, Solvent, Microscopy, Software