Quantification of VWF
Seventy-two hours after the transfection of HEK293T cells, the
supernatant medium was collected and was subsequently concentrated on
Amicon Centrifugal 50K filter devices (Millipore, USA) to one-fourth of
the original volume. Additionally, the transfected cells were lysed in
M-per reagent buffer (Thermo Scientific, USA). The amount of VWF antigen
(VWF:Ag) secreted into the medium and present in the lysate were
measured using a particle-based turbidimetric assay, and the mean values
(n=9) were expressed as a percentage of the corresponding wt.