An In-Gel Fluorescence Technique to Study Autophagy in Human Embryonic Kidney 293 Cells

Melissa Rogers; Riley Hatton; Robert Youker; Heather Coan

doi:10.21467/ajgr.17.1.1-10

Authors

Melissa Rogers Biology Department, College of Arts and Sciences, Western Carolina University
Riley Hatton Chemistry Department, College of Arts and Sciences, Western Carolina University https://orcid.org/0009-0004-2207-9968
Robert Youker Biology Department, College of Arts and Sciences, Western Carolina University https://orcid.org/0000-0002-3986-1655
Heather Coan Biology Department, College of Arts and Sciences, Western Carolina University https://orcid.org/0009-0000-3509-2185

DOI:

https://doi.org/10.21467/ajgr.17.1.1-10

Abstract

Autophagy is a recycling pathway used by cells to maintain homeostasis during times of stress. Common methods for the analysis of autophagy include western blotting and fluorescent imaging, but these methods are time consuming, expensive, and complex, making them inherently difficult for research laboratories at primarily undergraduate research institutions. We propose an in-gel fluorescence method for the analysis of autophagy in cells transfected with a dual-reporter plasmid. We verify that this method allows users to detect autophagic stimulation and is a cost effective, less complex method that sidesteps the challenges inherent with existing methods.

Keywords:

autophagy, in-gel fluorescence, LC3 I/II

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