Gene targeting strategy for B-hCDH17 mice. The extracellular region (except signal peptide) of mouse Cdh17 was replaced by human CDH17 extracellular region sequences in B-hCDH17 mice.

Strain specific analysis of CDH17 mRNA expression in wild-type C57BL/6 mice and B-hCDH17 mice by RT-PCR. Liver, colon and small intestine RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hCDH17 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human CDH17 primers. Human CDH17 mRNA was detectable in colon and small intestine of homozygous B-hCDH17 mice, not in liver. Meanwhile, the mouse Cdh17 mRNA was detectable in colon and small intestine of wild-type C57BL/6 mice, not in liver.

Simple Western analysis of CDH17 protein expression in homozygous B-hCDH17 mice. Liver, colon and small intestine lysates were collected from wild-type C57BL/6 (+/+) mice and homozygous B-hCDH17 mice (H/H), and then analyzed by Simple Western with species-specific anti-hCDH17 antibody (abcam, ab109190). 3ug total proteins were loaded for simple western analysis. Human CDH17 protein was detected in the colon and small intestine of homozygous B-hCDH17 mice, whereas human CDH17 protein was not detected in the liver of homozygous B-hCDH17 mice.

Representative human CDH17 expression in different tissues of B-hCDH17 mice by IHC. Tissues were stained with antibodies specific for human CDH17 (A-B, D-L) or anti-IgG antibody (C). Colon and ileum of B-hCDH17 mice show human CDH17 positive. Heart, liver, spleen, lung, kidney, stomach, and pancreas of B-hCDH17 mice show human CDH17 negative. Human gastric carcinoma cell line SNU-5 as a positive control (A); Colon tissues from the wild-type C57BL/6 mice as a negative control (B); Other tissues were got from homozygous B-hCDH17 mice (C-L). Original magnification ×200. Abbreviations: IHC, immunohistochemistry. The antibodies were provided by the client.