Colocalization Assay with Fluorescent-tagged ATG8 Using a Nicotiana benthamiana -based Transient System

Jinyan Mai; Dandan Shang; Faqiang Li; Na Luo

doi:10.21769/BioProtoc.4486

Colocalization Assay with Fluorescent-tagged ATG8 Using a Nicotiana benthamiana -based Transient System

Bio Protoc. 2022 Aug 20;12(16):e4486. doi: 10.21769/BioProtoc.4486.

Authors

Jinyan Mai¹, Dandan Shang¹, Faqiang Li², Na Luo¹

Affiliations

¹ College of Life Sciences, South China Agricultural University, Guangzhou 510642, China.
² College of Life Sciences, Guangdong Provincial Key Laboratory of Protein Function and Regulation in Agricultural Organisms, South China Agricultural University, Guangzhou 510642, China.

Abstract

Autophagy is an evolutionarily conserved intracellular degradation process. During autophagy, a set of autophagy-related (ATG) proteins orchestrate the formation of double-bound membrane vesicles called autophagosomes to engulf cytoplasmic material and deliver it to the vacuole for breakdown. Among ATG proteins, the ATG8 is the only one decorating mature autophagosomes and therefore is regarded as a bona fide autophagic marker; colocalization assays with ATG8 are wildly used as a reliable method to identify the components of autophagy machinery or autophagic substrates. Here, we describe a colocalization assay with fluorescent-tagged ATG8 using a tobacco ( Nicotiana benthamiana )-based transient expression system.

Keywords: ATG8; Autophagy; Colocalization assay; Nicotiana benthamiana; Transient expression.