FIG. 8.

Rta-dependent-DcR3 expression is mainly from its direct binding to the DcR3 promoter. (A) Serum-starved SW480 cells were transfected with GFP-, GFP-Rta-, or GFP-Rta-NLSm-expressing plasmids. The samples in lanes 1 to 3 were dimethyl sulfoxide solvent controls for the PI 3-K inhibitor (20 μM of LY294002)-treated samples in lanes 4 to 6. The Western blots were probed with antibodies of Rta (for GFP-Rta and GFP-Rta-NLSm), total Akt, phosphorylated Akt (p-Akt), DcR3, or the control cellular protein β-actin. A longer DcR3 exposure was provided to show the basal level of DcR3 expression under starvation conditions. (B) GFP-Rta, GFP-Rta-NLSm, or vector control plasmid (2 μg/10⁶ cells) was transfected with the luciferase reporter driven by the DcR3 promoter region, pDcR3 −1010, into SW480 cells. Luciferase activity was measured and is presented in relation to that of GFP alone. (Bottom) Equal expression of GFP-Rta and GFP-Rta-NLSm by Western analysis. β-Actin was a loading control.