Cells were detached using trypsin and pelleted, resuspended and washed
with 100 μL PBS 3 times. Cells were then incubated at 4 °C for
30 minutes with PAR1 mouse monoclonal antibody (Invitrogen), and washed 3 times
with PBS. Cells were then incubated with goat anti-mouse IgG-FITC antibody
(Invitrogen) for 30 minutes at 4 °C, and washed 3 times with PBS. Cells
were then fixed in 2% formaldehyde and cells were analyzed using a
BDFacs Canto II flow cytometer (BD Biosciences, San Jose, CA) equipped with a
488 nm argon laser and a 530 bandpass filter (FL1). A minimum of 10,000 events
were counted for each data point. The data was analyzed using the FACSDiva
version 6.1.1 software. Fluorescence data is expressed as mean arbitrary
fluorescence units and were gated to include all healthy cells.
with 100 μL PBS 3 times. Cells were then incubated at 4 °C for
30 minutes with PAR1 mouse monoclonal antibody (Invitrogen), and washed 3 times
with PBS. Cells were then incubated with goat anti-mouse IgG-FITC antibody
(Invitrogen) for 30 minutes at 4 °C, and washed 3 times with PBS. Cells
were then fixed in 2% formaldehyde and cells were analyzed using a
BDFacs Canto II flow cytometer (BD Biosciences, San Jose, CA) equipped with a
488 nm argon laser and a 530 bandpass filter (FL1). A minimum of 10,000 events
were counted for each data point. The data was analyzed using the FACSDiva
version 6.1.1 software. Fluorescence data is expressed as mean arbitrary
fluorescence units and were gated to include all healthy cells.