Fig. 3

GSK-3 inactivation requires long-term incubation with T-cells to enhance CTLs killing of tumors. GSK inhibitor increases cytolytic killing of CTLs when present during over the cell culture period needed to induce differentiation. CTLS were generated by incubating splenocytes from OT-1 Tg mice with OVA-peptide for 7 days. SB415286 was added to cultures on a day 0 or b day 6. On day 7 T-cells were washed and a 4 h cytolytic assay performed using EL-4 cells pulsed or non-pulsed with Ova-peptide as target cells.  c depicts T-cells only treated with SB415286 following the wash step and prior to the 4 h CTL assay (In panels a and b any residual SB415296 was washed away). Error bars based on triplicate values in individual experiments, data shown representative of 3 independent experiments. * P < 0.05; ** P < 0.005; *** P < 0.0005. d Figure shows examples of T-cells interacting and killing tumor targets (EL4-OVA cells labelled in red). The CTL then goes on to lyse the cell as can be seen from the characteristic bubbling of the cell cytoplasm and the clear vacuole. Data shown representative of three independent experiments