FIG. 2.

FKHR is mislocalized in PTEN-null cells. (A) Fluorescence microscopy of GFP-FKHR in PTEN-plus and PTEN-null cells. The indicated cells were transiently transfected, as described in Materials and Methods, with a plasmid encoding GFP-FKHR or GFP-FKHR;AAA. At 24 h after transfection, GFP-FKHR was detected by fluorescence microscopy in living cells. (B) Quantification of the results obtained with GFP-FKHR as shown in panel A. The percentage of cells with nuclear localization of GFP-FKHR was determined by manual counting of GFP-positive nuclei. Data shown are the mean and standard error of duplicate experiments and are representative of two independent experiments. (C) Localization of FKHRL1 in 786-O and ACHN cells. 786-O and ACHN cells were fractionated into cytoplasmic and nuclear compartments by hypotonic lysis and Dounce homogenization. Equivalent cell fractions were loaded on the gel and immunoblotted with anti-FKHRL1, anti-β-tubulin, and anti-pRB antibodies as indicated.