FIG 3.

Screening of actin-associated proteins suggests MCPyV ST expression induces filopodium formation. (A) HEK-293 cells were cotransfected with 1 μg of EGFP-mDia2 and empty control vector or cotransfected with 1 μg of EGFP-mDia2 and ST-Flag. Twenty-four hours later, the cells were fixed and permeabilized, and GFP fluorescence was analyzed by direct visualization; in addition, the cells were stained with rhodamine-phalloidin and a Flag-specific antibody. (B) HEK-293 cells were cotransfected with 1 μg of EGFP and IRSp53-myc or cotransfected with 1 μg of EGFP-ST and IRSp53-myc. Twenty-four hours later, the cells were fixed and permeabilized, and GFP fluorescence was analyzed by direct visualization; in addition, the cells were stained with rhodamine-phalloidin and a Myc-specific antibody. (C) HEK-293 cells were transfected with 1 μg of EGFP or EGFP-ST. Twenty-four hours later, the cells were fixed and permeabilized, and GFP fluorescence was analyzed by direct visualization; in addition, the cells were stained with rhodamine-phalloidin and a myosin X-specific antibody. The enlarged box shows myosin X staining at the tips of filopodia (arrows). All the slides were analyzed using a Zeiss LSM 700 confocal laser scanning microscope.