Cleavage of Bcl-2 in Jurkat cells treated with agonistic anti-Fas Ab or coincubated with tumor cells.
In (A), Fas-sensitive and Fas-resistant Jurkat cells 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 Bcl-2 expression (anti–Bcl-2 mAb 100, Santa Cruz). In (B), Fas-sensitive 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, using epithelial-specific anti-α6β4 mAb and magnetic beads. Negatively selected Jurkat cells were lysed and analyzed by Western blotting for expression of Bcl-2. Better separation between the Bcl-2 bands was observed in B than in A, owing to longer electrophoresis.
