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. 2008 Jul 1;99(2):294–304. doi: 10.1038/sj.bjc.6604459

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Copyright © 2008 Cancer Research UK

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Immunoblot analyses of candidate TRAIL signalling regulators. Immunoblotting was performed on lysates from the indicated glioma early passage cell lines or 293T cells transiently transfected with expression plasmids encoding the various apoptotic pathway components. (A–D) Anti-DR5 (A), anti-caspase-8 (B), anti-FADD (C) and anti-XIAP (D) signals were quantitated using a chemidoc instrument and plotted relative to the 293T transfectant lysates (set at 100%). A nonspecific band detected by the anti-DR5 antibody is indicated by an asterisk. Long exposures using autoradiograph film revealed some signals too weak to be detected by chemidoc (+). Illustrative immunoblots are inset within each graph. (E) Autoradiography was used to assay cFLIP_L and p53 expressions in early passage lines and 293T cells transfected with the cFLIP_L expression plasmid (cFLIP_L, GAPDH immunoblots) or empty vector (p53 immunoblot). Irrelevant lanes separating the 293T transfectant signals from those of the early passage lines have been removed.