mRNA expression analysis

Strain specific analysis of IL6 and IL6R gene expression in B-hIL6/hIL6R mice by RT-PCR. Mouse Il6 and Il6r mRNA were detectable in spleen of wild-type mice (+/+). Human IL6 was detectable only in homozygous B-hIL6/hIL6R mice (H/H) after stimulated with LPS for 3h. Human IL6R mRNA was detectable in homozygous B-hIL6/hIL6R mice but not in wild-type mice with or without LPS stimulation.

Protein expression analysis

Strain specific IL6R expression analysis in homozygous B-hIL6/hIL6R mice by flow cytometry. Splenocytes were collected from WT (+/+) and homozygous B-hIL6/hIL6R mice (H/H), and analyzed by flow cytometry with species-specific anti-IL6R antibody. Mouse IL6R was detectable in WT mice. Human IL6R was exclusively detectable in homozygous B-hIL6/hIL6R mice but not in WT mice.

IL6R expression pattern analysis by IHC

Protein expression analysis in plasma

Strain specific IL6R expression analysis in wild-type C57BL/6 mice and homozygous humanized B-hIL6/IL6R mice by ELISA. Plasma(EDTA) was collected from wild-type C57BL/6 mice (female, 6-week-old, n=3) and homozygous B-hIL6/IL6R mice (female, 6-week-old, n=3). Expression level of human IL6R were analyzed by ELISA (human IL6R ELISA kit: R&D, DR600). Human IL6R was exclusively detectable in homozygous B-hIL6/IL6R mice. Values are expressed as mean ± SEM.

Analysis of spleen leukocytes cell subpopulations in B-hIL6/hIL6R mice

Analysis of spleen T cell subpopulations in B-hIL6/hIL6R mice