siRNA-mediated silencing of human <i>CBLB</i> is effective and leads to hyper-responsiveness of CD8⁺ T cells.

<p>Human CD8⁺ T cells were nucleofected with <i>CBLB</i> siRNA #10 or control siRNA and stimulated with anti-CD3 and anti-CD28 as indicated. Similar results were obtained with non-overlapping siRNA oligonucleotide #11 (not shown). (A) Supernatants were taken two days post-transfection and the amount of IFNγ secreted was measured by BioPlex technology. Means ± SEM of two independent experiments are shown. p = 0.01 (CD8⁺<i>CBLB</i> siRNA versus CD8⁺ control siRNA, double stimulated). (B) Human CD8⁺ T cells were nucleofected with <i>CBLB</i> siRNA or control siRNA and stimulated with anti-CD3 and anti-CD28 in the presence or absence of TGFβ. Supernatants were taken two days post-transfection and the amount of IFNγ secreted was measured by BioPlex technology. IFNγ levels of each group without TGFβ were set 100%. Representative results of two independent experiments are shown. Silencing efficacy of <i>cblb</i> siRNA was analyzed by (A) immunoblotting (One experiment representative of three experiments is shown) or (C) qRT-PCR. Means ± SEM of two independent experiments are shown.