FIGURE 5.

Activation of caspases upon TNFα/CHX exposure. Apoptosis was induced by TNFα/CHX for the indicated times (0, 2, and 4 h) in HeLa cells in which FAM129B was knocked down by exposure for 72 h with siRNA (SC-92820-A) and in control cultures transfected with scrambled siRNA (control). A, the cell extracts were analyzed by Western blotting using antibodies directed against FAM129B and the mouse monoclonal antibody 1C12 that recognizes full-length human procaspase-8, the 41 and 43-kDa proteolytic intermediates and the 18-kDa subunit of the activated caspase. B, Western blotting of cell extracts using an antibody directed against intact and cleaved PARP and an antibody recognizing the 17 and 12-kDa subunits of activated caspase-3. The cell extracts in A and B were also immunoblotted with β-tubulin antibodies.