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. 2009 Sep 9;284(44):30159–30166. doi: 10.1074/jbc.M109.027995

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© 2009 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 3. — PTEN complex is enriched in the nucleus. A, WT-PTEN and NLS-PTEN were transfected into PC3 cells. Cell lysates were applied to a gel filtration column. Fractions were collected and subjected to immunoblotting (IB) with anti-PTEN antibody. The percentage of PTEN in the complex fractions was calculated by dividing the amount of PTEN in the high molecular mass fractions by total PTEN (complex + monomer). The amount of PTEN in these fractions was determined through densitometry analysis. B, NLS-PTEN was transfected into PC3 cells, the nuclear and cytoplasmic fractions were prepared and run through a gel filtration column, and fractions were collected and immunoblotted with anti-PTEN antibody. C, percentage of NLS-PTEN in the complex fractions was calculated for nuclear (Nuc) and cytosolic (Cyt) extracts as described in A and depicted in a bar graph. D, nuclear and cytosolic fractions (column input) were immunoblotted with anti-PTEN and anti-HDAC (nuclear marker) antibodies.