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-xL(
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.