Fig 8.

RIP, c-FLIP and PED/PEA-15 localize the DISC in non-rafts in TRAIL resistance. (A) The lipid raft and non-raft fractions from TRAIL-resistant LN443 cells were subjected to Western blotting of lipid raft markers caveolin-1 and Fyn. (B) Chemical analysis of cholesterol content in the lipid raft and non-raft fractions of LN443 cells. (C) LN443 cells were untreated (left panel) or treated with 300 ng/ml TRAIL for 15 min. (right panel) and lipid raft and non-raft fractions were separated and examined on Western blots for DR5, caspase-8 (Casp-8), FADD, RIP, c-FLIP and caveolin-1. (D) LN443 cells were transfected with RIP, c-FLIP and PED/PEA-15 siRNA, respectively, and then treated with 300 ng/ml TRAIL and subjected to discontinuous sucrose density gradients. In total, 2–10 fractions were examined by Western blot analysis of DR5 and caspase-8.