Ethanol decreases lipid raft colocalization of Lck and LAT in Jurkat cells. Jurkat cells were untreated (U), treated for 24 hours with 25 mM ethanol alone (E), 5 μg/ml phytohemagglutinin (PHA) alone for 5 minutes (PHA), or ethanol followed by PHA (E+PHA). Detergent-resistant lipid raft domains were isolated from Triton-X100 solubilized Jurkat cells using sucrose density gradient ultracentrifugation. Lipid raft colocalization of Lck and LAT, as well as raft-resident marker Flotillin, was analyzed by immunoblotting of raft (#2 to #4) and nonraft (#7 to #8) fractions using specific antibodies.