Differential effects of overexpression of Rab5 and Rab22 on autophagy in PC12 cells with or without NGF

Methods Mol Biol. 2015:1298:295-304. doi: 10.1007/978-1-4939-2569-8_25.

Abstract

Macroautophagy selectively recycles damaged or unneeded proteins and organelles by degradation via targeting to the autophagosome. The following method seeks to identify candidate Rab GTPases that likely modulate autophagy in PC12 cells during nerve growth factor (NGF) starvation. This microscopy-based assay is a single cell-based quantification of the presence of autophagosomes by fluorescently labeled markers in response to the overexpression of Rabs and mutants in the presence or absence of NGF.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Autophagy / drug effects*
  • Blotting, Western
  • Cricetinae
  • Gene Expression
  • Nerve Growth Factor / pharmacology*
  • PC12 Cells
  • Plasmids / genetics
  • Rats
  • Single-Cell Analysis / methods*
  • Transfection
  • rab5 GTP-Binding Proteins / genetics*

Substances

  • Nerve Growth Factor
  • rab5 GTP-Binding Proteins