Generation Strategy
Figure 1. Schematic representation of gene targeting at the murine Pd-l2 (mPd-l2) locus to generate the humanized PD-L2 (hPD-L2) mouse. A chimeric humanized Pd-L2 gene containing the human extracellular domain and the mouse intracellular Pd-l2 domain was inserted into the endogenous murine pd-l2 locus.
RT-PCR Genotyping of Humanized PD-L2 Mice
Figure 2. Genotyping of humanized PD-L2 mice by RT-PCR using primer set P1/P2 for wild type (WT) gene and primer set P3/P4 for humanized chimeric transgene (HE) were used to confirm knock-in at the targeted locus. PCR fragments of size 404 bp in WT mice and 405 bp in the humanized mice was observed and confirmed by sequencing (not shown). M:100bp Plus DNA ladder from Transgene (BM311)
Expression of hPD-L2 in Activated Splenocytes of Humanized PD-L2 Mice
Figure 3. Splenocytes from humanized PD-L2 mice were analyzed by flow cytometry after activation. The hPD-L2+ (but not mPD-L2+) cells were detectable in the homozygous (HO) hPD-L2 mice, while hPD-L2+ and mPD-L2+ cells were both detectable in heterozygous (HE) humanized PD-L2 mice.