Processing of caspase-3 in Jurkat cells treated with agonistic anti-Fas Ab or coincubated with tumor cells.
In (A) Jurkat cells, including Fas-resistant (Fas-R), Neo control, CrmA, and Bcl-2, were treated with CH-11 (200 ng/mL) for 16 hours. At the end of the incubation period, the cells were lysed and assessed by Western blotting for caspase-3 (anti-CPP32 mAb, 2.5 μg/mL). In (B) Fas-sensitive (Fas-S) and Fas-resistant Jurkat cells were coincubated with tumor cell (20:1 tumor-to-Jurkat cell ratio) for 16 hours. At the end of the coincubation period, Jurkat lymphocytes were negatively selected by removal of tumor cells, with the use of epithelial-specific anti-α6β4 mAb and magnetic beads. Negatively selected Jurkat cells were lysed and analyzed by Western blotting for processing of caspase-3.
