Targeting strategy
Gene targeting strategy for B-hS1PR1 mice. The exon 2 of mouse S1pr1 gene that encodes the full-length protein was replaced by human S1PR1 exon 2 in B-hS1PR1 mice.
Protein expression analysis in T, B and NK cells of spleen
Strain specific S1PR1 expression analysis in homozygous B-hS1PR1 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hS1PR1 mice (H/H), and analyzed by flow cytometry with species-specific anti-S1PR1 antibody. mS1PR1 and hS1PR1 were both detectable in T cells (A), B cells (B) and NK cells (C) of spleen from wild-type and homozygous mice, as anti-mS1pr1 antibody and anti-hS1PR1 antibody were both cross-reactive between human and mouse.
Protein expression analysis in monocytes, macrophages and neutrophils of spleen
Strain specific S1PR1 expression analysis in homozygous B-hS1PR1 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hS1PR1 mice (H/H), and analyzed by flow cytometry with species-specific anti-S1PR1 antibody. mS1PR1 and hS1PR1 was both detectable in monocytes (A), macrophages (B) and neutrophils (C) of spleen from wild-type and homozygous mice, as anti-mS1pr1 antibody and anti-hS1PR1 antibody were both cross-reactive between human and mouse.
Protein expression analysis in T, B and NK cells of lymph nodes
Strain specific S1PR1 expression analysis in homozygous B-hS1PR1 mice by flow cytometry. Lymph nodes cells were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hS1PR1 mice (H/H), and analyzed by flow cytometry with species-specific anti-S1PR1 antibody. mS1PR1 and hS1PR1 was both detectable in T cells (A), B cells (B) and NK cells (C) of lymph nodes from wild-type and homozygous mice, as anti-mS1pr1 antibody and anti-hS1PR1 antibody were both cross-reactive between human and mouse.
mRNA expression analysis
Strain specific analysis of S1PR1 mRNA expression in wild-type C57BL/6 mice and B-hS1PR1 mice by RT-PCR. Spleen RNA were isolated from wildtype C57BL/6 mice (+/+) and homozygous B-hS1PR1 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human S1PR1 primers. Mouse S1pr1 mRNA was detectable only in wild-type C57BL/6 mice. Human S1PR1 mRNA was detectable only in homozygous B-hS1PR1 mice but not in wild-type mice.
* When publishing results obtained using this animal model, please acknowledge the source as follows: The animal model [B-hS1PR1 mice] (Cat# 111029) was purchased from Biocytogen.
