Figure 4.

Apoptosis and BAD interaction with 14-3-3 and Bcl-X_L in JEG3 cells. Apoptosis was assessed (A; n = 3) by immunocytochemical detection of cyt 18 filament neoepitopes in transfected JEG3 cells in the presence or absence of EGF in a FiO₂ = 20% or hypoxia (FiO₂ < 1%). *, P < 0.05 vs. no EGF, t test. Cell lysates from JEG3 cells ± 100 ng/ml EGF, transfected with either wild-type GFP-BAD (B) or GFP-mBADS112A (C), were immunoprecipitated with an antibody to BAD before immunoblotting for 14-3-3 proteins and Bcl-X_L (27 kDa). The upper band of the 14-3-3 proteins detected showed interaction with BAD in the wild-type transfectants (B), but this interaction was not detected in the mutant BAD transfectants (C). The other band detected by the 14-3-3 antibody showed interaction with BAD in both transfectants.