Schematic presentation of the third generation lentiviral vector used in the transgenesis studies and Southern blot image of a copy number analysis. (A) CMV indicates cytomegalovirus promoter; Ψ, packaging signal; cPPT, central polypurine tract. The TG cassette contains the human phosphoglycerate kinase (hPGK) promoter, human VEGF-D cDNA truncated from C- and N-termini (hVEGF-D), and woodchuck hepatitis virus PRE element (wPRE). SIN indicates that the lentiviral vector is self-inactivating because of the deletion in U3 LTR to form ΔU3; U5, 5′ LTR; RRE, rev-responsive element. (B) Analysis of the 4 hVEGF-D TG founder mice (1, 2, 3, and 4) and 3 F₁ mice (2a, 2b, and 2c) from founder number 2. The copy numbers of standards (left half of the image) are indicated (copies/15 μg of mouse tail genomic DNA). wPRE was used as a probe sequence. Molecular weight marker (MW) between the standard and sample lanes is 2.5 kb. A vertical line has been inserted to indicate a repositioned gel lane; 1, 1 to 5 copies; 2, 7 to 10 copies; 3, 5 to 7 copies; 2a, 5 to 7 copies; 2b, 7 to 10 copies; and 2c, 7 to 10 copies.