Fig. 4. BID activation and cytochrome c release during PKC/Ras-mediated apoptosis. In A, Jurkat, Jurkat/ras, Jurkat/FADDm/ras, or Jurkat/Fasm/ras cells were left untreated or treated with high-dose PMA for 24 h, and lysates were prepared. Equal amounts of total protein from each cell lysate were separated on a SDS-PAGE gel, transferred to nitrocellulose, and immunoblotted with an anti-BID Ab. In B, the cytosol fraction from untreated cells or treated cells was prepared. Samples containing equal amounts of cytosol proteins were separated on a 15% SDS-PAGE gel, and immunoblot analysis of cytochrome c (Cyt. C) was performed using an anti-cytochrome c Ab.