Pedigree of the family with the inherited bleeding disorder and thrombin generation in the affected relatives. (A) Pedigree of the family with the inherited bleeding disorder. Affected individuals are indicated by shapes filled half-way; unaffected individuals are indicated by nonfilled shapes; and nonfilled shapes with ? indicate disease status unknown. Blood samples were collected from all living relatives to assess coagulation parameters and ultimately to perform whole exome sequencing. (B-G) Thrombin generation in the affected relatives. Thrombin generation was initiated with (B) 1 or (C) 5 pM tissue factor in normal pooled plasma (NP), the proband (II:2), her son (III:1), and husband (II:1). Thrombin generation could be restored in the affected relatives by addition of blocking antibodies against TFPI or protein S. Thrombin generation was initiated with 1 pM tissue factor in (D) normal pooled plasma (NP), (F) the proband (II:2), (G) her son (III:1), and (E) husband (II:1) in the absence or presence of blocking antibodies against TFPI or protein S. Similar results were obtained with 5 pM tissue factor (data not shown).