Phosphorylation of eIF3f by CDK11p46 is important for the inhibition of translation and induction of apoptosis. (A) A375 cells were co-transfected with pcDNA3, or pcDNA3-eIF3f, or pcDNA3-eIF3fSATA and pGL3-SV40 carrying the luciferase reporter gene. At 24 h post-transfection, cells were lysed and luciferase activity was measured by a luminometer. (B) Real-time RT-PCR was performed as described. Untransfected A375 cells and reactions without reverse transcription served as controls. (C) Apoptosis was measured by Acridine Orange/Ethidium Bromide staining. (D) (E) Luciferase mRNA was translated in an in vitro translation system in the presence of buffer, GST, or indicated recombinant proteins. Synthesized luciferase activity was measured with a luminometer. (F) A375 cells were transfected with indicated plasmids. Caspase 3/7 activity was measured 48h after transfection.