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. 2014 May 6;9(5):e95248. doi: 10.1371/journal.pone.0095248

Figure 3. CD8+ T cells require direct prolonged contact with target cells to kill KC.

Figure 3

Primary KC loaded with ovalbumin peptide were co-cultured with CD8+ T cells from EGFP+OT-1 mice and imaged over a 30-hour period in the presence of a cell-permeable indicator dye that fluoresces red upon activation of caspase-3. A. The number of caspase-3 positive keratinocytes in each 5 hour time period per experiment was determined by standardized objective analysis of movies by specialized imaging software. N = 6, error bars are SEM. B. In primary KC culture, contaminating fibroblasts are readily distinguished by their morphology. These also uptake peptide and present as targets for CTL. KC and fibroblast survival in co-culture with OT-1 cells; (Pooled results N = 5, error bars are SD). C. Addition of caspase-3 inhibitor, Z-DVED-FMK, an inhibitor of caspase-3, resulted in abrogation of CTL-mediated apoptosis. (Pooled results N = 5, error bars are SD). D. T cell death, following a similar pattern to KC death, is abrogated by the addition of Z-DVED-FMK (Pooled results N = 3, error bars are SD).