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Fig. 7. cFLIPL overexpression in keratinocytes potently inhibits TRAIL-mediated apoptosis. Stably transfected keratinocytes were generated by retroviral infection of either PINCO-control vector or PINCO-cFLIPL in HaCaT keratinocytes as described in Materials and Methods. Overexpression of cFLIPL was confirmed using 50 µg protein of total cellular lysates by western blotting and subsequent analysis for cFLIPL. As control, cell lysates were blotted against anti-procaspase-8 antibody (C15) (A). (B) For functional analysis, cells were seeded in 96-well-plates in duplicates and were stimulated with the indicated concentrations of recombinant TRAIL for 16 hours. The percentage of living cells was normalized to mock-stimulated cells as marked on the y-axis. One of three representative experiments is shown.





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