Transient expression of the C-terminal domain in HeLa cells induces mitotic arrest. ( A ) HeLa cells were cotransfected with pEGFP-F and various pCMV-FLAG-Plk constructs and fixed as described in Materials and Methods . DNA profiles of GFP-positive cells were determined by fluorescence-activated cell sorter (FACS) analysis. Left (ungated) and Right (GFP-positive) represent the DNA profiles from total cell populations and GFP-positive transfected cells after gating, respectively. 1, DNA profiles from transfectants with pCMV-FLAG vector; 2, with pCMV-FLAG-Plk K82M; 3, with pCMV-FLAG-Plk WT; 4, with pCMV-FLAG-Plk T210D; 5, with pCMV-FLAG-ΔC1–401; 6, with pCMV-FLAG-C306–603. ( B ) The percent of G 2 /M phase cells determined from the GFP-positive population by ModFit analysis. Data represent three independent experiments. ( C ) Western blot analysis of expressed proteins from transfected cell lysates with anti-FLAG antibody.
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