The murine platelet and plasma factor V pools are biosynthetically distinct and sufficient for minimal hemostasis by Hongmin Sun, Tony L. Yang, Angela Yang, Xixi Wang, and David Ginsburg Blood Volume 102(8):2856-2861 October 15, 2003 ©2003 by American Society of Hematology

The construction of AlbFv or Pf4Fv BACs The construction of AlbFv or Pf4Fv BACs. (A) The transgene cassette used for homologous recombination into the Alb or Pf4 BACs. The locations of PCR and RT-PCR primers A to D are illustrated by arrows. The construction of AlbFv or Pf4Fv BACs. (A) The transgene cassette used for homologous recombination into the Alb or Pf4 BACs. The locations of PCR and RT-PCR primers A to D are illustrated by arrows. The cassette contains the murine Fv cDNA with a chimeric 5′ intron and a 3′ UTR deletion tag, followed by an internal ribosome entry site sequence (IRES) preceding the enhanced green fluorescent protein (EGFP) coding sequence and a bacterial kanamycin resistant gene (KAN). (B) Homologous recombination between the flanking sequences of the constructs and corresponding BACs. Exons 1-12 of the murine Alb gene and exons 1-3 of murine Pf4 gene were replaced with an FV cDNA minigene. Hongmin Sun et al. Blood 2003;102:2856-2861 ©2003 by American Society of Hematology

Expression patterns of Fv transgenes. Expression patterns of Fv transgenes. (A) RT-PCR for Fv transgene expression was performed on tissue samples from 3 Pf4 Fv (Pf4A-C) and 6 Alb Fv (AlbA-F) transgenic mice. (B) RT-PCR for Fv transgene expression was performed on tissue samples from AlbA and Pf4A transgenic rescued FV-null mice (Tg+Fv–/–). Hongmin Sun et al. Blood 2003;102:2856-2861 ©2003 by American Society of Hematology

Tail vein bleeding time. Tail vein bleeding time. Tail vein bleeding times of Pf4A and AlbB mice Tg+ are shown in seconds. Rescue mice (Tg+Fv–/–) are compared with control littermates carrying the transgene and at least one wild-type Fv allele (Fv+ = Fv+/– or Fv+/+). The mean and SD are also indicated for each genotype. Hongmin Sun et al. Blood 2003;102:2856-2861 ©2003 by American Society of Hematology

Levels of Fv transgene expression. Levels of Fv transgene expression. The level of Fv expression observed in the plasma (A) and platelet (B) pools of the transgenic Fv-null rescued mice (Tg+Fv–/–) are shown here for all 6 albumin Fv BAC lines and one Pf4 Fv BAC line. The FV level observed in platelets or plasma of a control Fv+/– mouse, corresponding to one endogenous copy of the normal Fv gene, is arbitrarily defined as 50%. SEs are indicated. Hongmin Sun et al. Blood 2003;102:2856-2861 ©2003 by American Society of Hematology