Blood, Vol. 115, Issue 22, 4580-4587, June 3, 2010

The mutation N528S in the von Willebrand factor (VWF) propeptide causes defective multimerization and storage of VWF
Blood Haberichter et al. 115: 4580

Supplemental materials for: Haberichter et al

Files in this Data Supplement:

• Figure S1. Heterozygous co-expression of N528S-VWF and wild-type VWF (JPG, 327 KB) -
  (A) HEK293 cells were transfected with varying amounts of N528S-VWF and wild-type-VWF as listed on the x-axis. The concentration of VWF in the conditioned media was assessed by VWF ELISA. (B) HEK293 cells were transfected at a N528S-VWF:wild-type-VWF ratio of 3:1. Cells were immunostained using monoclonal AVW-5 that does not recognize wild-type-VWF-myc-his (red) and rabbit-anti-myc that does not recognize the N528S-VWF lacking a myc-his tag (green); colocalization of both proteins is shown in yellow (arrow). (C) HEK293 cells were transfected with varying amounts of N528S-VWF and wild-type-VWF-myc-his as listed on the x-axis. Differential antibody recognition (see B above) was used to visualize multimers of coexpressed N528S-VWF (red) and wild-type-VWF-myc-his (green) with colocalization in yellow. The majority of secreted VWF was expressed from the wild-type allele and very little N528S-VWF was detected.