Fig. 6.

Presentation of a TAP-downregulation-induced peptide in human tumors treated with Nucl-TAP. a Downregulation of TAP mRNA in human tumor cells incubated with Nucl-TAP. Cells were treated with Nucl-siRNAs and 24 h later mRNA was isolated and quantified by qRT-PCR. Data represent means and SEM performed in triplicates (n = 2). b and c Downregulation of HLA-A, B, and C analyzed by flow cytometry at different times as indicated (n = −3). b Data represent means and SEM performed in quadruplicates. c Representative experimental well at 96 h. d and e Presentation of a TAP downregulation-induced peptide in TAP-deficient (TAP KO) or TAP-sufficient 518A2 and MZ18.51 tumor cells treated with Nucl-siRNAs for 72 h, and cultured with a CD8⁺ T cell clone that recognized the HLA-A2-TAP downregulation-induced peptide complex²⁹. d In vitro cytotoxicity after 4 h coculture was determine by lactate dehydrogenase assay. e INFγ production after 20 h stimulation was measured by ELISA. Means and SEM of quadruplicate wells (n = 1). Statistical analyses using one-way ANOVA test and Dunnet posttest for comparison between untreated and treated cells. Differences are indicated: ****P < 0.001, ***P < 0.005, **P < 0.01, *P < 0.05