Fig. 7.

Extracellular regulated kinase (ERK) phosphorylation was decreased in Nck1 knock-down Jurkat cells. Jurkat T cells were treated with specific Nck1 short interfering (si)RNA (Nck1) or control siRNA (Ctl). After 48 h of cultivation, cells were subsequently stimulated with 10 µg/ml anti-CD3ε monoclonal antibody (mAb) plus 10 µg/ml anti-CD28 mAb or with 6 µg/ml phytohaemagglutinin (PHA) plus 1 ng/ml phorbol myristate acetate (PMA) or none (−) for 10 min at 37°C. Cytoplasmic proteins were immunoblotted with anti-phospho-Erk1/2 (Thr202/Tyr204, Thr185/Tyr187) antibody.