Journal: Cells

Article Title: Coagulation Factor Xa Induces Proinflammatory Responses in Cardiac Fibroblasts via Activation of Protease-Activated Receptor-1

doi: 10.3390/cells10112958

Figure Lengend Snippet: Effect of thrombin and FXa on mRNA expression of F2r (PAR-1 gene) ( A , B ) and F2rl1 (PAR-2 gene) ( C , D ). Rat atrial ( n = 5–9) and ventricular ( n = 6–8) CFs were stimulated with 100 nM of FXa for 4 and 24 h. Gene expression of F2r and F2rl1 was measured by RT-qPCR. Data were normalized to the housekeeping gene Cyclophilin-A. Results are expressed as median bars, with dots indicating individual rat CF isolations. Statistical analysis was performed using the non-parametric Wilcoxon test for paired samples (* p < 0.05, ** p < 0.01).

Article Snippet: Human CFs (200 cells/cm 2 ) were seeded in 96-well plates, cultured for 24 h, and serum-starved for 24 h. Subsequently, cells were incubated with human FXa (100 nM) with or without PAR-1 antagonist (1 µM, SCH79797, Tocris, Bioscience, Bristol, UK).

Techniques: Expressing, Quantitative RT-PCR

Journal: Cells

Article Title: Coagulation Factor Xa Induces Proinflammatory Responses in Cardiac Fibroblasts via Activation of Protease-Activated Receptor-1

doi: 10.3390/cells10112958

Figure Lengend Snippet: Effect of PARs silencing on FXa-mediated CCL2 and IL6 expression in human CFs ( A , B ; n = 8–9). Before exposure to FXa (100 nM), human atrial CFs were transfected with 10 μM of either scrambled F2R (PAR-1) or F2RL1 (PAR-2) specific siRNA and incubated for 48 h. Gene expression was measured by RT-qPCR after 4 h of incubation with FXa. Data were normalized to the housekeeping gene Cyclophilin-A. Statistical analysis was performed using the non-parametric Friedman test for paired samples ( CCL2 : overall p < 0.001; IL6 : overall p < 0.001) and Dunn’s multiple comparison test (** p < 0.01, **** p < 0.0001). Effect of silencing on F2R (PAR-1) and F2Rl1 (PAR-2) gene expression ( C , D ; n = 14). Data were analysed using the non-parametric Wilcoxon test for paired samples (*** p < 0.001). Effect of FXa inhibition by rivaroxaban on CCL2 and IL6 expression in human CFs ( E , F ; n = 4). Statistical analysis was performed using the non-parametric Friedman test for paired samples ( CCL2: overall p < 0.001; IL6: overall p < 0.001) and Dunn’s multiple comparison test (* p < 0.05). Results are expressed as median bars, with dots indicating individual human CF isolations.

Article Snippet: Human CFs (200 cells/cm 2 ) were seeded in 96-well plates, cultured for 24 h, and serum-starved for 24 h. Subsequently, cells were incubated with human FXa (100 nM) with or without PAR-1 antagonist (1 µM, SCH79797, Tocris, Bioscience, Bristol, UK).

Techniques: Expressing, Transfection, Incubation, Quantitative RT-PCR, Comparison, Inhibition

Journal: Cells

Article Title: Coagulation Factor Xa Induces Proinflammatory Responses in Cardiac Fibroblasts via Activation of Protease-Activated Receptor-1

doi: 10.3390/cells10112958

Figure Lengend Snippet: Effect of PARs silencing on CCL2 and IL6 expression ( A , B ; n = 6). Human atrial CFs were transfected with 10 μM of either scrambled PAR-1 or PAR-2 specific siRNA before exposure to TRAP-14 (100 μM). Gene expression was measured by RT-qPCR after 4 h of incubation. Data were normalized to the housekeeping gene Cyclophilin-A. Results are expressed as median bars, with dots indicating individual human CF isolations. Statistical analysis was performed using the non-parametric Friedman test ( CCL2 : overall p < 0.0001; IL6 : overall p < 0.0001) test for paired samples and Dunn’s multiple comparison test (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Human CFs (200 cells/cm 2 ) were seeded in 96-well plates, cultured for 24 h, and serum-starved for 24 h. Subsequently, cells were incubated with human FXa (100 nM) with or without PAR-1 antagonist (1 µM, SCH79797, Tocris, Bioscience, Bristol, UK).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Incubation, Comparison

Journal: Cells

Article Title: Coagulation Factor Xa Induces Proinflammatory Responses in Cardiac Fibroblasts via Activation of Protease-Activated Receptor-1

doi: 10.3390/cells10112958

Figure Lengend Snippet: Proliferation assay on human atrial CFs ( n = 3). Cells were incubated with 100 nM FXa with and without SCH79797 (1 μM). Proliferation was measured 24 h after exposure to the stimuli via BrdU colorimetric assay. Results are expressed as fold-change relative to CTRL (mean bars). Statistical analysis was performed using the repeated measures one-way ANOVA ( p < 0.01) and Šidák’s multiple comparison test (** p < 0.01). Lines depict paired data points.

Article Snippet: Human CFs (200 cells/cm 2 ) were seeded in 96-well plates, cultured for 24 h, and serum-starved for 24 h. Subsequently, cells were incubated with human FXa (100 nM) with or without PAR-1 antagonist (1 µM, SCH79797, Tocris, Bioscience, Bristol, UK).

Techniques: Proliferation Assay, Incubation, Colorimetric Assay, Comparison