Figure 1.

Genetic and expression analysis of 2bIbαLV-transduced bone marrow transplantation (BMT) recipients. (a) PCR analysis of BMT recipients shows the presence of transgene in recipients. Genomic DNA was prepared from primary (1°) and secondary (2°) 2bIbα lentiviral vector (LV) transduced hematopoietic stem cells (HSC) recipients. GPIbα^null and C57BL/6J wild-type mice were used as controls. 2bIbα LV plasmid DNA was used as a positive control for human GPIbα (hGPIbα). Absence of mGPIbα PCR product confirmed the GPIbα^null background. The Vwf gene was used as an internal control. PCR product sizes; hGPIbα (458 bp), mGPIbα (486 bp), and Vwf (727 bp). (b) Immunofluorescent staining of mouse platelets. Platelets were isolated from GPIbα^null mice that received 2bF8 LV-transduced HSC (upper panel) and untransduced GPIbα^null control mice (lower panel) and stained for hGPIbα (green) and murine VWF (red). Nonspecific isotype-matched primary antibodies were used to assess background staining (data not shown). Bar = 10 µm.