NRASQ61K is required for survival of the CTCL cell line Hut78. (A) siRNA against NRAS (siNRAS1 and siNRAS2) were transfected in SeAx, Hut78, MyLa, and HH cell lines by the Amaxa kit and protocol. At 72 hours after transfection, cells were lysed and lysates subjected to Western blot. Efficiency of knockdown was measured by anti-NRAS antibodies. Levels of phosphorylation of MEK and ERK were assessed by anti-phospho-specific MEK and anti-phospho-specific ERK in SeAx and Hut78 cells. Equal loading was verified by antitubulin antibodies. (B) Apoptosis was measured starting 3 days after siRNA transfection for 24, 48, and 72 hours. Specific apoptosis was calculated as described in “Cell death assays.” Data are representative of 3 independent experiments.
