hTNFSF11(2)

Fig.1 Detection of TNFSF11 expression in thymus by RT-PCR. Tissue RNA was extracted from wild-type C57BL/6 litters (WT) and homozygous hTNFSF11(2) knockin mice (HO) (7 weeks old), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mRNA primers.  Mouse Tnfsf11 mRNA (167 bp) was detectable only in wild-type C57BL/6 mice. Human TNFSF11 mRNA (295 bp) was detectable only in homozygous hTNFSF11(2) knockin mice but not in wild-type mice.

Fig.2 Detection of RANKL (TNFSF11) expression on CD4+ T cells in spleen of WT C57BL/6 mice and HO hTNFSF11(2) mice by FACS. hRANKL was exclusively detectable in hTNFSF11(2) humanized mice but not in wild-type mice. hRANKL was exclusively detectable on CD4+ T cells in hTNFSF11(2) humanized mice, whereas mRANKL was exclusively detectable on CD4+ T cells in wild-type C57BL/6 mice. CD4+T cells were collected from spleen of wild-type C57BL/6 mice (male, 19 weeks old) and homozygous hTNFSF11(2) humanized mice (male, 19 weeks old), stimulated with anti-mCD3e (5μg/mL), anti-mCD28 (5μg/mL) and mouse IL-2 (100ng) in vitro for 3 days, and analyzed by flow cytometry with anti-mRANKL and anti-hRANKL. 

Abbr. HO, homozygous; WT, wild type.