Studying the Role of AMPK in Autophagy

Methods Mol Biol. 2018:1732:373-391. doi: 10.1007/978-1-4939-7598-3_24.

Abstract

AMPK is an energy-sensing kinase and is required for the induction and progression of the autophagy process. In this chapter, we describe experimental approaches to study the steady state and flux of autophagy in response to AMPK activation. For this purpose, we provide detailed protocols for the measurement of general as well as AMPK-specific autophagy markers by immunoblot and immunofluorescence analysis.

Keywords: A-769662; AICAR; AMP-activated protein kinase; Autophagy; Bafilomycin A1; Flux analysis; Glucose deprivation; Immunoblot; Immunofluorescence; LC3.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AMP-Activated Protein Kinases / metabolism*
  • Aminoimidazole Carboxamide / analogs & derivatives
  • Aminoimidazole Carboxamide / pharmacology
  • Animals
  • Autophagy / drug effects*
  • Biomarkers / analysis
  • Biphenyl Compounds
  • Cell Line, Tumor
  • Enzyme Activation / drug effects
  • Enzyme Activators / pharmacology*
  • Enzyme Inhibitors / pharmacology*
  • Fibroblasts
  • Fluorescent Antibody Technique / instrumentation
  • Fluorescent Antibody Technique / methods
  • Humans
  • Immunoblotting / instrumentation
  • Immunoblotting / methods
  • Macrolides / pharmacology
  • Mice
  • NIH 3T3 Cells
  • Pyrones / pharmacology
  • Ribonucleotides / pharmacology
  • Thiophenes / pharmacology

Substances

  • Biomarkers
  • Biphenyl Compounds
  • Enzyme Activators
  • Enzyme Inhibitors
  • Macrolides
  • Pyrones
  • Ribonucleotides
  • Thiophenes
  • Aminoimidazole Carboxamide
  • bafilomycin A1
  • AMP-Activated Protein Kinases
  • AICA ribonucleotide
  • 4-hydroxy-3-(4-(2-hydroxyphenyl)phenyl)-6-oxo-7H-thieno(2,3-b)pyridine-5-carbonitrile