Citation Details

Role of Bcl-2 family proteins in a non-apoptotic programmed cell death dependent on autophagy genes

Authors	Shimizu S, Kanaseki T, Mizushima N, Mizuta T, Arakawa-Kobayashi S, Thompson CB, Tsujimoto Y
In	Nat Cell Biol (2004) 6(12): 1221-1228
Genes of Interest	Bax Bak Bcl-2 Bcl-xL LC3 Beclin 1 APG5
Cells used in publication	Embryonic fibroblast (MEF), mouse    Mouse embryonic fibroblasts (MEF)    Connective Tissue Cells    Primary Cells    Species: mouse    Tissue Origin: embryo
Substrate	Plasmid (general) siRNA
Applications	RNAi Co-Transfection
Topics	Apoptosis Microscopy of nucleofected cells Signal transduction
Research Area	Cancer Research/Cell Biology
Relevant Products	Nucleofector Device MEF Starter Nucleofector Kit MEF Nucleofector Kit 1 MEF Nucleofector Kit 2
Reporter Gene	GFP unspecified
Vector Backbone	pUC

Research Field

Programmed cell death can be divided into several categories including type I (apoptosis) and type II (autophagic death). The Bcl-2 family of proteins not only regulates apoptosis, but also controls non-apoptotic programmed cell death that depends on the autophagy genes.

Nucleofection Experiments

Primary and SV40 T antigen-transformed wild-type and Bax/Bak–/– mouse embryonic fibroblasts (MEFs), as well as APG5+/+ and APG5−/− MEFs were transfected and/or cotransfected with plasmids expressing human Bax, human Bak, human Bcl-2, human Bcl-xL, or GFP–LC3.
The cells were also transfected with siRNAs to mouse Bcl-x, mouse Bcl-2, mouse Bax, mouse Beclin 1, or mouse APG5. In the case of transfection with complementary DNA and siRNA, cDNAs were transfected, and after 24 h siRNAs were introduced.

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