hOSM

Fig.1 Detection of OSM expression in spleen by RT-PCR. Tissue RNA was extracted from wild-type C57BL/6 litters (WT) and homozygous hOSM knockin mice (HO) (n=2, female, 10 weeks old), then cDNA libraries were synthesized by reverse transcription, followed by PCR with human or mouse specific mRNA primers. Mouse Osm mRNA (360 bp) was detectable only in wild-type C57BL/6 mice. Human OSM mRNA (274 bp) was detectable only in homozygous hOSM knockin mice but not in wild-type mice.

Fig.2 Detection of OSM expression from splenocytes by ELISA. Splenocytes were isolated from female 10-week-old wild-type (WT) and homozygous hOSM knock-in (HO) mice, then seeded in 12-well plates at a density of 2×106 cells/well in 2 mL culture medium. Following in vitro stimulation with 5 μg/mL anti-mouse CD3 antibody and 5 μg/mL anti-mouse CD28 antibody for 24 or 48 h, human OSM levels in the culture supernatants were measured using a human OSM ELISA kit. Results showed that hOSM was detectable exclusively in HO mice. The expression of OSM significantly increased in response to anti-CD3/CD28 stimulation in HO mice. 

Abbr. N.D., not detected; M, DNA marker; HO, homozygous; WT, wild type.