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 | RNAiCo-Transfection |
| Topics | ApoptosisMicroscopy of nucleofected cellsSignal transduction |
| Research Area | Cancer Research/Cell Biology |
| Relevant Products |  Nucleofector Device MEF Starter Nucleofector KitMEF 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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