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Fig. 3. The nuclear localization of endogenous mSTI1 and mSTI1-EGFP is enhanced by G1/S-phase arrest. (A) Untransfected NIH3T3 fibroblasts (left), NIH3T3 fibroblasts stably transfected with pB-EGFP (middle) or with pB-mSTI1-EGFP (right) were exposed to hydroxyurea (10 mM) for 10 hours. Untransfected cells were immunostained to detect endogenous mSTI1 with SF1 polyclonal rabbit anti-mSTI1 and Cy3-conjugated donkey anti-rabbit antibodies, and transfected cells were fixed in 3.7% paraformaldehyde before mounting and visualization by confocal laser fluorescence microscopy. Scale bars, 10 µm. (B) Cells were exposed to hydroxyurea as above, and cells demonstrating cytoplasmic fluorescence or nuclear and cytoplasmic fluorescence were quantified. The bars represent standard deviations.
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