Figure 6, TAP-independent PMEL_209-217 presentation requires endocytic recycling but not Hrd1-dependent ERAD.

(A) Various amounts of a buf1280(A2; GFP↓) control lysate were loaded on an SDS-PAGE gel together with 40 μl of a lysate derived from Hrd1-silenced buf1280(A2; Hrd1↓) clones #1.22 and #1.23 (shRNA construct #1) (left panel). CTL7 degranulation measured in response to buf1280(A2) cell lines stably transduced with Hrd1-specific shRNA construct #1 (buf1280(A2; Hrd1↓) #1.22 and buf1280(A2; Hrd1↓) #1.23) or a GFP-specific control shRNA construct (buf1280(A2; GFP↓). Error bars represent the standard deviation from the mean of three independent experiments. A One-way ANOVA with Dunnett’s post test was performed for statistical evaluation (right panel). (B, C) CTL7 degranulation 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. Both experiments were repeated three times. Error bars represent the standard deviation from the mean of these three independent experiments. A One-way ANOVA with Dunnett’s post test was performed for statistical evaluation (NS = not significant). Some data shown in Fig. 6C was also included in Fig. 3A, 4D, and 5D.