Species specific SIRPα and CD47 expression analysis in B-hSIRPA/hCD47,Prkdc/Il2rg KO mice by flow cytometry. Splenocytes isolated from C57BL/6 (+/+) mice and homozygous B-hSIRPA/hCD47,Prkdc/Il2rg KO mice (H/H) were analyzed by flow cytometry with anti-SIRPα and anti-CD47 antibodies. Human SIRPα and CD47 were exclusively detectable in homozygous B-hSIRPA/hCD47,Prkdc/Il2rg KO mice but not in C57BL/6 mice. Mouse SIRPα was detectable in C57BL/6 and homozygous B-hSIRPA/hCD47,Prkdc/Il2rg KO mice, indicating that this anti-mouse SIRPα antibody was cross-reacting with human SIRPα. 

Species specific SIRPα and CD47 expression analysis in B-hSIRPA/hCD47,Prkdc/Il2rg KO mice by flow cytometry. Splenocytes were isolated from C57BL/6 (+/+) mice and homozygous B-hSIRPA/hCD47,Prkdc/Il2rg KO mice (H/H) stimulated with anti-mCD3ε in vivo and were analyzed by flow cytometry with anti-SIRPα and anti-CD47 antibodies. Human SIRPα and CD47 were exclusively detectable in homozygous B-hSIRPA/hCD47,Prkdc/Il2rg KO mice but not in C57BL/6 mice. Mouse SIRPα was detectable in C57BL/6 mice and homozygous B-hSIRPA/hCD47,Prkdc/Il2rg KO mice, indicating that this anti-mouse SIRPα antibody was cross-reacting with human SIRPα. 

Analysis of leukocyte subpopulations in spleen by FACS

Splenocytes were isolated from C57BL/6, B-NDG mice and B-hSIRPA/hCD47,Prkdc/Il2rg KO mice (n=3, 11-week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. Single live cells were gated for CD45 population and used for further analysis as indicated here. T, B and NK cells were undetectable in B-NDG mice and B-hSIRPA/hCD47,Prkdc/Il2rg KO mice.