hCDCP1(2)

Fig.1 Detection of CDCP1 expression in colon by RT-PCR. Colon RNA was extracted from male 6-week-old WT mice (n=2) and heterozygous hCDCP1(2) knockin mice (n=4), then cDNA libraries were synthesized by reverse transcription, followed by PCR with specific mRNA primers. Mouse Cdcp1 mRNA (479 bp) in both wild-type C57BL/6 mice and heterozygous hCDCP1(2) knockin mice. Human CDCP1 mRNA (634 bp) was detectable only in heterozygous hCDCP1(2) knockin mice but not in wild-type mice. 

Abbr. M, marker; HE, heterozygous; WT, wild type.

Fig.2 Detection of CDCP1 expression in skin and colon by qPCR. Skin and colon RNA were harvested from male 6-week-old WT mice (n=2) and heterozygous hCDCP1(2) knockin mice (n=4), then cDNA libraries were synthesized by reverse transcription, followed by qPCR with h/m Cdcp1 and mouse Gapdh primers. Relative expression represents the h/m Cdcp1 mRNA level relative to its average expression in WT mice. (Mean±SEM, T-test, ns, no significance).

Abbr. HE, heterozygous; WT, wild type.

Fig.3 Detection of CDCP1 expression in hCDCP1 knockin mice by WB. Tissues lysates were collected from male 6-week-old WT mice (n=2) and heterozygous hCDCP1 mice (n=4), and then analyzed by western blot with anti-CDCP1 antibody. 20 μg of total proteins were loaded for western blotting analysis. The positive control used are HCT116 (PC1) and HT29 (PC2) cells, and the negative control used is MCF-7 cells. The full length CDCP1 protein (~140KD) is detectable in skin, heart, colon and stomach tissues in both WT mice and heterozygous hCDCP1(2) mice, as the antibody is cross-reactive between human and mouse CDCP1. The cleaved CDCP1 fragment (~70KD) is not detactable in normal tissues from both WT mice and heterozygous hCDCP1(2) mice. 

Abbr. M, marker; HE, heterozygous; WT, wild type; PC, positive control; NC, negative control.