Figure 1.
Pedigree of the family with bleeding disorder.
Squares and circles indicate males and females, respectively, and arrow indicates the proband. Black color denotes affected individuals. A slash through the symbol indicates decreased individuals.
Table 1.
Main laboratory findings of the patient and family members.
Figure 2.
Analysis of plasma and platelet VWF multimers.
Plasma (A) and platelet (B) VWF multimers were assessed by 1.6% SDS-agarose gel electrophoresis and Western blotting. Normal plasma samples were diluted 1∶20 and patient plasma samples were diluted 1∶5. Samples from family members and a normal control (N) were indicated. Platelet samples were not available from IV-4.
Figure 3.
Identification of a VWF gene mutation.
Sequence analysis of the VWF gene in the proband detected a 6-bp nucleotides deletion in exon 28. The mutation caused D1529V1530 deletion (ΔD1529V1530) in VWF A2 domain. The ADAMTS13 cleavage site is indicated by an arrow.
Table 2.
Expression of WT and mutant VWF in transfected HEK293 cells.
Figure 4.
Multimer analysis of recombinant VWF expressed in HEK293 cells.
Multimeric analysis of recombinant VWF was performed with the conditioned medium from HEK293 cells transfected with plasmids for WT and the mutant, individually or in combination. Samples from the vector-transfected cells were used as a negative control.
Figure 5.
Detection of WT and mutant VWF in transfected HEK293 cells by immunostaining.
HEK293 cells were transfected with a control vector or plasmids expressing WT or mutant VWF. The cells were stained for VWF (red), PDI (green) or DAPI (nucleus, blue). In the right column panels, merged pictures of green, red and blue channels are shown. The images were obtained with a confocal microscope. Scale bars: 10 µm.