Figure 3

Evidence for enhanced cell survival in vitro . FETα and FETα-DN cells were grown to 80% confluence followed by growth factor deprivation for 48 h and probed for differences in apoptosis as reflected by several approaches. (A) Western blot analysis was performed for PARP and cleaved PARP antibodies, using actin as a loading control. (B) DNA fragmentation assay was utilized to determine apoptosis. Error bars represent S.E. (C) Western blot analysis was performed for AKT phosphorylation at S473, using total AKT as the experimental control. (D) DNA fragmentation assay of LY294002 (25uM) treated FETα cells under GFDS conditions described in the Material and Methods. Error bars represent S.E. FETα cells were grown to 80% confluence followed by deprivation of growth factors in the absence or presence of 5 ng/ml TGFβ for 48 h. Assessment of apoptotic activity of exogenous TGFβ was determined (E) Western blot analysis probing for XIAP and survivin, using actin as a loading control. (F) DNA fragmentation assay. Error bars represent S.E.