Fig. 3.
VWF-cleaving protease activity in lysates and conditioned media of cells transiently expressing ADAMTS13.
Diluted lysates and conditioned media from transfected HEK 293 cells were activated with BaCl2 and incubated with 2.5 μg VWF. NHP dilutions were used for assay calibration. Multimeric analysis of VWF was carried out by SDS gel electrophoresis in 1% agarose gels, and VWF was detected by immunostaining. Sample dilutions are indicated. VWF-cp assay was carried out with cell lysates (lanes 6-8) and conditioned medium (lanes 10-12) from transfections with plasmid pCMV/VWF-cp and the control vector (lanes 5 and 9) and with NHP (lanes 1-3) and buffer (lane 4) as probes.
