Figure 5

2-DG-mediated up-regulation of TRAIL-R2 is independent of p53 and CHOP. A, Mel-RM and MM200 cells were transfected with the control or p53 siRNA. Twenty-four hours later, whole cell lysates were subjected to Western blot analysis. B, Mel-RM and MM200 cells were transfected with the control or p53 siRNA. Twenty-four hours later, cells were treated with 2-DG (10 μM) for a further 16 hours. Upper panel: The cell surface expression of TRAIL-R2 was measured using flow cytometry. The data in y axes represent mean fluorescence intensity (MFI). Lower panel: Total RNA was isolated and subjected to Real-time PCR analysis for TRAIL-R2 mRNA expression. The relative abundance of mRNA expression before treatment was arbitrarily designated as 1. C, Whole cell lysates from Mel-RM and MM200 cells transduced with the control or CHOP shRNA were subjected to Western blot analysis. The arrow head points to non-specific bands generated by the antibody against CHOP. Note that the cells were treated with TM (3 μM) for 16 hours before harvest to better visualize CHOP. D, Mel-RM and MM200 cells transduced with the control or CHOP shRNA were treated with 2-DG (10 μM) for 16 hours. Left panel: The cell surface expression of TRAIL-R2 was measured using flow cytometry. The data in the y axes represent mean fluorescence intensity (MFI). Right panel: Total RNA was isolated and subjected to Real-time PCR analysis for TRAIL-R2 mRNA expression. The relative abundance of mRNA expression before treatment was arbitrarily designated as 1. The data shown are either the mean ± SE (B & D), or representative (A & C), of three individual experiments.