Cell Survival and Apoptosis Assays for E1B 19K and Bcl-2 Family Members

Programmed cell death, or apoptosis, is an important event in the normal development and homeostasis of multicellular organisms (1 ). Apoptosis is regulated by a number of genes that promote or suppress cell death. The bcl-2 family of genes constitute important regulators of apoptosis. Several members of the bcl-2 family such as bcl-2 (2 ) and bcl-x _L (3 ) suppress apoptosis, whereas certain other members such as bax (4 ), bak (5 ), and bik (6 ) promote apoptosis Certain animal viruses such as adenovirus, Epstein-Barr virus, herpesvirus simairi, and African swine fever virus also encode structural and functional homologs of Bcl-2. The methods currently available to determine the anti- or pro-apoptotic activities of various cellular and viral Bcl-2 homologs are cumbersome. Here, we describe highly reproducible, simple methods that we have developed to assay the activities of adenovirus E1B 19 kDa, Bcl-2, and various pro-apoptotic interacting partners.