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Methods in enzymology

Kinetic analysis of autophagosome formation and turnover in primary mouse macrophages.


PMID 19200894

Abstract

Macrophages enlist autophagy to combat infection by a variety of bacteria, viruses, and parasites. In response to this selective pressure, some pathogenic microbes have acquired strategies to evade or tolerate autophagy. Accordingly, infected cells may accumulate numerous autophagic vacuoles/autophagosomes when microbial products either stimulate their formation or inhibit their maturation. To distinguish between the two mechanisms, we describe methods to assess the impact of infection on the kinetics and amplitude of autophagosome formation and maturation within mouse macrophages by microscopy or Western analysis using antibodies specific for endogenous or recombinant LC3 protein.