Western Blot Protocols for Analysis of CCN Proteins and Fragments in Exosomes, Vesicle-Free Fractions, and Cells

Kisho Ono; Yuka Okusha; Manh Tien Tran; Koki Umemori; Takanori Eguchi

doi:10.1007/978-1-0716-2744-0_5

Western Blot Protocols for Analysis of CCN Proteins and Fragments in Exosomes, Vesicle-Free Fractions, and Cells

Methods Mol Biol. 2023:2582:39-57. doi: 10.1007/978-1-0716-2744-0_5.

Authors

Kisho Ono¹, Yuka Okusha², Manh Tien Tran³, Koki Umemori¹, Takanori Eguchi⁴

Affiliations

¹ Department of Oral and Maxillofacial Surgery, Okayama University Hospital/Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan.
² Division of Molecular and Cellular Biology, Department of Radiation Oncology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA.
³ Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, Japan.
⁴ Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, Japan. eguchi@okayama-u.ac.jp.

PMID: 36370343
DOI: 10.1007/978-1-0716-2744-0_5

Abstract

Cellular Communication Network (CCN) proteins are growth factors that play key roles in many pathophysiological events, including bone formation, wound healing, and cancer. CCN factors and fragments generated by metalloproteinases-dependent cleavage are often associated with extracellular matrix (ECM) or small extracellular vesicles (sEVs) such as exosomes or matrix-coated vesicles. We provide reliable methods and protocols for Western blotting to analyze CCN factors and fragments in cells, sEVs, and vesicle-free fractions.

Keywords: CCN family proteins; CCN fragments; CTGF; Cell communication network; Exosomes; Extracellular matrix; Extracellular vesicles; Matricellular proteins; Matrix vesicles; Metalloproteinases.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Blotting, Western
CCN Intercellular Signaling Proteins / metabolism
Cell Communication
Exosomes* / metabolism
Extracellular Matrix / metabolism
Extracellular Vesicles* / metabolism

Substances

CCN Intercellular Signaling Proteins