Figure 2, Cytosolic processing of PMEL_209-217.

(A) CTL7 activation measured by IFNγ ELISA in response to TAP-sufficient (blue bars) or TAP-inhibited (white bars) target cells (NT, not tested). (B, C) CTL7 degranulation measured in response to the indicated cell lines is shown as in Fig. 1B. CTL7 activation (black bars) and HLA-A2 surface levels on target cells (white bars) are depicted in bar diagrams. The experiment in Fig. 2C was repeated three times. Error bars represent the standard deviation from the mean of these three independent experiments. A paired two-tailed t-test was performed for statistical evaluation. Surface-HLA-A2 levels were measured in only two of these three experiments (white bars). (D, E) CTL7 activation measured by IFNγ ELISA (dark blue bars). The experiment in Fig. 2E was repeated four times. Error bars represent the standard deviation from the mean of these four independent experiments. A One-way ANOVA with Dunnett’s post test was performed for statistical evaluation. Surface-HLA-A2 levels were measured by flow cytometry in only two of these four experiments (white bars). Relatively low T cell recognition of T2(∆SS) cells in Fig. 2D reflects consistently lower sensitivity of the IFNγ ELISA compared to LAMP degranulation (Fig. 2C). (F) T cell activation of clone CTL7 (black bars), tyrosinase_1-9-specific clone CTL 210/9 (red bars) and tyrosinase_369-377-specific clone CTL IVSB (green bars) measured using the degranulation assay in response to the indicated cell lines. HLA-A2 surface levels are shown in white bars. (G) Expression of melanocyte differentiation antigens analyzed by RT-PCR. The dashed line indicates where an irrelevant lane was removed. (H) CTL7 activation measured in response to the indicated cell lines or cell line mixtures. Degranulation is quantified in the rightmost panel. Error bars represent the standard deviation from the mean of two independent experiments.