hMADCAM1

Fig.1 Detection of MADCAM1 expression in spleen by RT-PCR. Spleen RNA was extracted from 8-week-old male wild-type C57BL/6 (WT) and homozygous hMADCAM1 knockin mice (HO), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mRNA primers. Mouse Madcam1 mRNA (349 bp) was detectable only in wild-type C57BL/6 mice. Human MADCAM1 mRNA (515 bp) was detectable only in homozygous hMADCAM1 knockin mice but not in wild-type mice. 

Abbr. M, marker; HO, homozygous; WT, wild type.

Fig.2 Detection of MADCAM1 expression in spleen by qPCR. Spleen RNA was extracted from 8-year-old male wild-type C57BL/6 (WT) (n=2) and homozygous hMADCAM1 knockin mice (HO) (n=2), then cDNA libraries were synthesized by reverse transcription, followed by qPCR with h/m Madcam1 and mouse Gapdh primers. Relative expression represents the h/m Madcam1 mRNA level relative to its average expression in WT mice. 

Madcam1 mRNA levels exhibited a significant increase in homozyguos hMADCAM1 knockin mice compared to WT mice by qPCR with human/mouse-shared primers.

Abbr. M, marker; HO, homozygous; WT, wild type.

Fig.3 Detection of human MADCAM1 expression in hMADCAM1 knockin mice by IHC. Tissues were collected from 8-week-old male WT C57BL/6 mice and HO hMADCAM1 knockin mice, then analyzed by IHC with anti-mouse MADCAM1 antibody. Black arrows indicate positive staining (brown). The scales are same in each image. Magnification, 20X. 

Mouse MADCAM1 was exclusively detectable in endothelial cells of mesenteric lymph nodes, small intestine and colon in WT mice but not in homozygous hMADCAM1 knockin mice.

Fig.4 Detection of human MADCAM1 expression in hMADCAM1 knockin mice by IHC. Tissues were collected from 8-week-old male WT C57BL/6 mice and HO hMADCAM1 knockin mice, then analyzed by IHC with anti-human MADCAM1 antibody. Black arrows indicate positive staining (brown). The scales are same in each image. Magnification, 20X.

Human MADCAM1 was obviously detectable in endothelial cells of mesenteric lymph nodes, small intestine and colon in homozygous hMADCAM1 knockin mice.

Abbr. HO, homozygous; WT, wild type.