A high-throughput protocol for monitoring starvation-induced autophagy in real time in mouse embryonic fibroblasts

Abstract

Summary: Autophagy measurement has been challenging due to the transient nature of autophagy vesicles, in which degradation of cargo occurs. Here, we present a protocol to monitor starvation-induced autophagy using a live high-throughput microscopy system in a fast and automated manner without the need for sample preparation. We provide a detailed protocol describing the generation of turboGFP-LC3B expressing mouse embryonic fibroblasts (MEFs), the measurement of autophagy over time and the analysis of data.For complete details on the use and execution of this protocol, please refer to Nowosad et al. (2020, 2021).

Keywords

• Cell Biology
• Cell-based Assays
• High Throughput Screening
• Microscopy