Fig 1: MASP-1 and platelet activation. Whole blood supplemented with corn trypsin inhibitor and acetylsalicylic acid was re-calcified and incubated with rMASP-1cf (10 μg/ml and 50 μg/ml) and fixed at different time points. The entire platelet population was identified with an anti-CD41 antibody conjugated with APC. The activated population was identified with an anti-CD62P antibody conjugated with BV421. (A) Addition of rMASP-1cf increased the percentage of platelets expressing CD62P (shown as mean with SD) in a time- and dose-dependent manner. Statistical analysis: The Shapiro-Wilk test confirmed normal distribution, and p-values for differences between groups were determined with an ANOVA test (ns not significant, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001). Number of experiments: Control: n=8; 10 μg/ml MASP-1: n=3; 50 μg/ml MASP-1: n=6). (B) The effect of 50 μg/ml rMASP-1cf was tested in the presence of PAR4 inhibitor BMS986120 and/or hirudin. The samples were fixed after 15min. Addition of the PAR4 inhibitor significantly reduced the effect of rMASP-1cf, while addition of hirudin canceled it out completely. Statistical analysis: The Shapiro-Wilk test confirmed normal distribution, and p-values for differences between groups were determined with an ANOVA test (ns not significant, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001). Number of experiments: Control: n=8; MASP-1 only: n=6; MASP-1+ BMS: n=3; MASP-1+ Hirudin: n=4; MASP-1+ BMS + Hirudin: n=4.
Fig 2: Trauma patients had significantly increased bleeding times in a microfluidic model of vessel injury.A The microfluidic model comprises a native lumen, injury site, and extravascular space. Streamlines from simulated flow are shown. Healthy controls form a clot at the injury site resulting in microfluidic hemostasis and generation of a bleeding time (BT). B Representative control microfluidic hemostasis timelapse. Transmitted light (top) and platelet (anti-CD41) fluorescence (bottom, pink). C Left, samples from trauma patients had longer BT compared to healthy control samples at both venous and arterial shear rates when perfused through the microfluidic devices. Right, while 100% of the healthy control samples clotted, trauma patient samples had reduced closure frequency compared to healthy controls. Bleeding time are shown as individual values with bars representing mean and error bars representing standard deviation. Closure frequency is shown as absolute value. ns: not significant, * p<0.05, ** p<0.01, *** p<0.001.
Fig 3: Despite similar amounts of clot deposited, trauma patient samples have massively reduced platelet contribution to microfluidic clot.A Trauma patient samples had slightly reduced clot surface area deposition but overall deposited substantial amount of clot at the injury site. B Trauma patient samples had massively reduced platelet deposition compared to healthy controls at both venous and arterial shear. C Representative trauma microfluidic hemostasis timelapse from a sample that achieved hemostasis. Transmitted light (top) and platelet (anti-CD41) fluorescence (bottom, pink). D Maximum platelet MFI fold change was significantly reduced in trauma patient samples. Traces are shown as mean lines with standard deviation indicated as error bars below. E, F Lag time and growth rate of total surface area deposition were not different between trauma and controls. Individual data points are shown with bars at mean and error bars indicating standard deviation. ns: not significant * p<0.05, ** p<0.01, *** p<0.001. MFI: mean fluorescence intensity.
Supplier Page from Novus Biologicals, a Bio-Techne Brand for Integrin alpha 2b/CD41 Antibody (M148) - BSA Free
Available conjugates: Available conjugates: UnconjugatedSizes Available: 0.1 mg (also 0.025 mg)