Figure 3.
Hypoxia induced apoptosis of TILs through EMAP-II in vitro. Jurkat cells cocultured with tumour cells. FITC-labelled annexin-V assay (A) and caspase-3 activity (B) were performed to determine the level of apoptosis. (i) Jurkats alone; (ii) Jurkats cocultured with tumour cells; (iii) Jurkats cocultured with cells pretreated with TNF-α/IFN-γ; (iv) Jurkats cocultured with cells pretreated with TNF-α/IFN-γ in the presence of R2B2 blocking antibodies; (v) Jurkats cocultured with cells pretreated with TNF-α/IFN-γ in the presence of control IgG. The data shown are the averages of three experiments. The data shown are the averages of three experiments (mean±s.e.m.). ** Indicates significance at P<0.05 in comparison to controls. (C) Concentration-dependent increase in caspase-3 activity in jurkats with decreasing concentrations of anti-EMAP-II antibody. Data are means; error bars represent s.e.m. (D) Caspase-3 activity in Jurkats over time. 1, Jurkats+HT29 pretreated with TNF-α/IFN-γ in normoxia; 2, Jurkats+HT29 pretreated with TNF-α/IFN-γ in hypoxia; 3, Jurkats+HT29 pretreated with TNF-α/IFN-γ and R2B2 antibodies in normoxia; 4, Jurkats+HT29 pretreated with TNF-α/IFN-γ and R2B2 antibodies in hypoxia. Data are presented as the mean±s.e.m. of at least three separate experiments.