Basic Information

Strain name
C57BL/6N-Mfap5tm1(MFAP5)Bcgen/Bcgen
Common name
B-hMFAP5 mice
Background
C57BL/6N
Catalog number
112885
Aliases
AAT9, MAGP-2, MAGP2, MFAP-5, MP25
NCBI gene ID
8076

Description

  • MFAP5 is the gene that encodes the human MAGP-2 protein. Microfibril associated glycoproteins 1 and 2 (MAGP-1, MAGP-2) are the protein components of extracellular matrix microfibril. These proteins interact with fibrin, the core component of microfibril, affecting the function of vertebrate microfibril. MAGPs regulates Notch signaling in conjunction with TGFβ and BMP. MAGP-2 has also been shown to be an important biomarker for several human cancers. MFAP5 is basically not expressed in tumor tissues, but only expressed in tumors of about 10% of endometrial cancer patients. However, MFAP5 expression is elevated in many tumor-associated fibroblasts (CAFs), including non-small cell lung cancer, pancreatic cancer, ovarian cancer, prostate cancer, and breast cancer.
  • Protein expression analysis: Human MFAP5 was detectable in heart, uterus, ovary, bladder and fat of homozygous B-hMFAP5 mice, but not in wild-type mice.
  • mRNA expression analysis: Mouse Mfap5 was only detectable in uterus of wild-type C57BL/6 mice. Human MFAP5 was detectable in uterus of homozygous B-hMFAP5 mice.

Targeting strategy

Gene targeting strategy for B-hMFAP5 mice.

The exons 4~10 of mouse Mfap5 gene that encode the functional region, including 3”UTR were replaced by human MFAP5 exons 4~10 in B-hMFAP5 mice. The promoter, 5’UTR region and signal peptide of the mouse gene are retained. The human MFAP5 expression is driven by endogenous mouse Mfap5 promoter, while mouse Mfap5 gene transcription and translation will be disrupted.

mRNA expression analysis

Strain specific analysis of MFAP5 mRNA expression in wild-type C57BL/6 mice and B-hMFAP5 mice by RT-PCR. Uterus RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hMFAP5 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human MFAP5 primers. Mouse Mfap5 mRNA was detectable in wild-type mice. Human MFAP5 mRNA was detectable only in homozygous B-hMFAP5 mice, but not in wild-type mice.

Protein expression analysis

Western blot analysis of MFAP5 protein expression in homozygous B-hMFAP5 mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hMFAP5 mice (H/H), and then analyzed by western blot with species-specific anti-MFAP5 antibody (Abcam, ab203828). 40 μg total proteins were loaded for western blotting analysis. MFAP5 was detected in heart, uterus, ovary, bladder and fat of homozygous B-hMFAP5 mice, but not in wild-type mice.

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