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Rab GTPases pp 295-304 | Cite as

Differential Effects of Overexpression of Rab5 and Rab22 on Autophagy in PC12 Cells with or without NGF

Part of the Methods in Molecular Biology book series (MIMB, volume 1298)

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.

Key words

Autophagy LC3B Rab GTPase GTP-binding protein NGF PC12 cells 

Notes

Acknowledgment

The authors’ research program is supported by the NIH/NIGMS grant R01 GM074692 (to G.L.).

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of Biochemistry and Molecular Biology, Peggy and Charles Stephenson Cancer CenterUniversity of Oklahoma Health Sciences CenterOklahoma CityUSA
  2. 2.Department of Biochemistry and Molecular Biology, Peggy and Charles Stephenson Cancer CenterUniversity of Oklahoma Health Sciences CenterOklahoma CityUSA

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