Generation and characterization of transgenic FcγRIIA/hPF4 mice.

(A) PCR detection of transgenic mice by means of human-specific primers for hPF4 and FcγRIIA, and control mouse-specific primers for whey acidic protein (WAP). Lane 1, transgenic hPF4; lane 2, transgenic FcγRIIA; lane 3, double-transgenic progeny; lane 4, wild-type mice; lane 5, human genomic DNA. (B) Western blot of platelet proteins separated on a 10% Nu-PAGE gel, transferred to PVDF, and immunostained with RTO, a human PF4-specific mouse monoclonal antibody, shows an intensely stained band at approximately 8 kd, consistent with the molecular weight of PF4. Lane 1, wild-type mice; lane 2, wild-type plus added recombinant hPF4; lane 3, transgenic hPF4 mice; lane 4, human platelet proteins; lane 5, recombinant hPF4. The lane with wild-type mouse platelet proteins does not show staining in the 8-kd region. The other stained bands at molecular weights of 55 and 25 kd represent immunostaining of immunoglobin heavy and light chains that react with the anti–mouse immunoglobulin secondary antibody.