title: Differential effects of overexpression of Rab5 and Rab22 on autophagy in PC12 cells with or without NGF.
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.
authors: Department of Biochemistry and Molecular Biology, Peggy and Charles Stephenson Cancer Center, University of Oklahoma Health Sciences Center, 975 NE 10th Street, BRC 419, Oklahoma City, OK, 73104, USA.
authors: M Caleb
authors: MC
authors: Marlin
authors: Guangpu
authors: G
authors: Li
mesh_terms: D000818:Animals; D001343:Autophagy / Q000187:drug effects*; D015153:Blotting, Western; D006224:Cricetinae; D015870:Gene Expression; D020932:Nerve Growth Factor / Q000494:pharmacology*; D016716:PC12 Cells; D010957:Plasmids / Q000235:genetics; D051381:Rats; D059010:Single-Cell Analysis / Q000379:methods*; D014162:Transfection; D020696:rab5 GTP-Binding Proteins / Q000235:genetics*
chemical_list: D020932:Nerve Growth Factor; D020696:rab5 GTP-Binding Proteins