Figure 1.

Genotyping and phenotyping of P-selectin and Rag2 double null mutants. (A) Genomic DNA from mouse tail clips were used as templates for PCR genotype screening for Rag2 allele and P-selectin allele. The blackened region represents the neomycin A resistance gene that was targeted into both wild-type alleles; the arrows represent the two forward primers and the single reverse primer used in each reaction. Each PCR contained the three primers. F1 represents the double heterozygote from the first cross between Rag2 −/− and P-selectin −/− mice. (B) Peripheral blood from F1 mice and double null mutants was labeled with anti-CD4-phycoerythrin or anti-CD8-fluorescein isothiocyanate and analyzed by using a Becton Dickinson FACScan flow cytometer. (C) Frozen lung sections were probed with biotinylated anti-mouse P-selectin antibody and followed with streptavidin-peroxidase. The enzyme conjugate was detected with metal-enhanced diaminobenzidine substrate. The arrows indicate lung endothelium.