Document Type

Book Chapter or Section

Publication Date

2019

Comments

This article has been peer reviewed. It is the authors' final version prior to publication in The Extracellular Matrix, Methods in Molecular Biology.

Volume 1952, 2019, Pages 157-191.

The final publication is available at Springer via http://dx.doi.org/10.1007/978-1-4939-9133-4_14.

Abstract

A growing body of research demonstrates modulation of autophagy by a variety of matrix constituents, including decorin, endorepellin, and endostatin. These matrix proteins are both pro-autophagic and anti-angiogenic. Here, we detail a series of methods to monitor matrix-induced autophagy and its concurrent effects on angiogenesis. We first discuss cloning and purifying proteoglycan fragment and core proteins in the laboratory and review relevant techniques spanning from cell culture to treatment with these purified proteoglycans in vitro and ex vivo. Further, we cover protocols in monitoring autophagic progression via morphological and microscopic characterization, biochemical western blot analysis, and signaling pathway investigation. Downstream angiogenic effects using in vivo approaches are then discussed using wild-type mice and the GFP-LC3 transgenic mouse model. Finally, we explore matrix-induced mitophagy via monitoring changes in mitochondrial DNA and permeability.

PubMed ID

30825174

Language

English

Available for download on Sunday, March 01, 2020

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