FIGURE 9.

Recombinant FAM129B restores the resistance to apoptosis. HeLa cells were co-transfected with FAM129B siRNA, the EGFP vector or recombinant EGFP-FAM129B for 48 h using Lipofectamine 2000. Apoptosis was then induced by incubation with TNFα/CHX for an additional 4 h. Immunoblotting using EGFP-specific antibodies showed that recombinant EGFP-FAM129B (lanes 2 and 4) and as a control, the EGFP protein (EGFP vector) (lanes 1 and 3) were both highly expressed in cells treated with TNFα/CHX and in untreated cells. Expression of the recombinant protein had no effect on the level of PARP or caspase-3 in the untreated controls (lane 2) compared with cells transfected with only the vector (lane 1). Following induction of apoptosis, the cells transfected with EGFP vector (lane 3) exhibited increased PARP cleavage and caspase-3 activation, whereas these effects were diminished in cells transfected with recombinant FAM129B (lane 4). β-Tubulin served as a loading control.