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Subcellular fractionation analysis. Cos7 cells were transfected with the indicated plasmids. Whole-cell lysate (W), cytoplasmic (C), and nuclear (N) fractions were prepared as described in Materials and Methods. Then 20 mg of protein from each fraction was subjected to Western blot analysis by using the indicated Abs. In all of the extracts prepared, Sp1 was present exclusively in the nuclear fractions, which demonstrated the accuracy of the fractionation (Lower).
