FIGURE 3.

HB9-GFP locus identification. (A) Schematic of the HB9-GFP transgene, showing positions of primers used for TLA. Primer sets were designed within the Mxn1 and the GFP sequences. (B) Genome-wide TLA coverage using Mnx1 primers. Peak at Chromosome 5 shows endogenous Mnx1 (denoted by black circle) and peak at Chromosome 15 shows inserted sequence (red circle). (C) Genome-wide TLA coverage using GFP primers, showing a peak at Chromosome 15 (red circle). (D) Regional coverage of the HB9-GFP insertion site on Chromosome 15, spanning 74 kb surrounding the integration site. (E) Schematic of the inserted sequence. Multiple copies of the transgene were inserted 3′ to 5′. HB9-GFP is located between Cdh6 and Cdh9 on Chromosome 15. Primers were designed to confirm the left and right junctions of the transgene with Chromosome 15. PCR product from homozygous HB9-GFP, heterozygous HB9-GFP, and C57BL/6J animals is denoted as “Tg/Tg,” “Tg/+,” “WT” respectively. (F) Confirmed insertion of the transgene with primers specific to the junction between the 3′ end of the transgene and Chromosome 15 (Primers 1 and 2). (G) Confirmed insertion of the transgene with primers specific to the 5′ end of the transgene and Chromosome 15 (Primers 3 and 4).