Fig. 5.
Measurement of the cytotoxic activity of PBMC against full-length, truncated TF-overexpressing and control HT29 cells. A truncated form of the EGFP-TF was expressed by mutating the Ser240 into a stop codon. HT29 cells transfected with pEGFP-TFwt and pEGFP-TFtrunc transfected and un-transfected HT29 cells (5×104 per well) were labelled with SNARF-1 in culture medium (200 μl final volumes). Ten random views within each well were recorded using a fluorescence microscope. The cells were then challenged with preparations of isolated PBM cells at a ratio of 10:1 (PBMC: HT29). After 4 h incubation at 37°C, ten random sectors were analysed as before. Analysis was carried out using the Image Pro plus program and the data were calculated as the percentage reduction in cell number after incubation with PBMC cells against the initial number of viable HT29 cells. The data represent the mean of duplicates±standard deviation