Strain specific FOLR1 and FOLR2 expression analysis in wild-type mice and homozygous B-hFOLR1/hFOLR2 mice by flow RT-PCR. Mouse Folr1 and Folr2 mRNA were detectable in splenocytes of wild-type (+/+) mice. Human FOLR1 and FOLR2 mRNA were detectable only in homozygous B-hFOLR1/hFOLR2 mice, but not in wild-type (+/+) mice. 

mRNA expression analysis

Strain specific FOLR1 expression analysis in wild-type mice and homozygous B-hFOLR1/hFOLR2 mice by flow cytometry. Mouse Folr1 mRNA was detectable in splenocytes of wild-type (+/+) mice. Human FOLR1 mRNA were only detectable in lung and liver of homozygous B-hFOLR1/hFOLR2 mice, but not in wild-type (+/+) mice.

Protein expression analysis

Strain specific FOLR1 expression analysis in wild-type mice and homozygous B-hFOLR1/hFOLR2 mice by western blot. Kidney, spleen and brain were collected from C57BL/6 mice (+/+) and homozygous B-hFOLR1/hFOLR2 mice (H/H), and analyzed by western blot with anti-FOLR1 antibody. FOLR1 was detectable in kidney and brain but not spleen from wild-type mice and homozygous B-hFOLR1/hFOLR2 mice, as the antibody is crossly reactive with human and mouse FOLR1.

Strain specific FOLR1 expression analysis in wild-type mice and homozygous B-hFOLR1/hFOLR2 mice by western blot. Kidney, spleen and brain were collected from C57BL/6 mice (+/+) and homozygous B-hFOLR1/hFOLR2 mice (H/H), and analyzed by western blot with anti-FOLR1 antibody. FOLR1 was detectable in liver, lung and fallopian tube from wild-type mice and homozygous B-hFOLR1/hFOLR2 mice, as the antibody is crossly reactive with human and mouse FOLR1.

Protein expression analysis in Bone marrow

Analysis of blood T cell subpopulations by FACS. Blood were isolated from female C57BL/6 and homozygous B-hFOLR1/hFOLR2 mice (n=3, 8-week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for CD3+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD8+ T cells, CD4+ T cells, and Tregs in homozygous B-hFOLR1/hFOLR2 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hFOLR1/hFOLR2 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in blood. Values are expressed as mean ± SEM.