Fig. 5. Characterization of the nucleo-cytoplasmic distribution of TIAR and TIAR mutants by cell fractionation. Cos-7 cells were transfected with the indicated TIAR and TIAR mutants fused with the FLAG epitope. Where indicated, cells were then treated with actinomycin D (5 µg/ml, 3 hours) before cell fractionation. Cytoplasmic (C) and nuclear (N) extracts were analyzed by western blot using anti-FLAG and anti-FBP to control the fractionation procedure.