Fig. 6. WIP binding to actin is essential for PDGF-induced ruffle formation. (A)
GST pull-down assay. The indicated GST-WIP constructs (5 µg per 10 µl
beads) were incubated with 0.05 µM G-actin. After SDS-PAGE, samples were
blotted with anti-actin monoclonal antibody. (B) Fluorescence micrographs of
TRITC-phalloidin-stained Swiss 3T3 fibroblasts transfected with control
plasmid (3T3pcDNA), WIP coding sequence (3T3pcDNA-WIP) or the deletion mutant
WIP
43-54 (3T3pcDNA-WIP43-54), before and after PDGF challenge.
Cells were grown on glass coverslips, serum starved for 3 days in the presence
of 0.5% FBS, stimulated or not with PDGF (50 ng ml-1) for 8 minutes
and then fixed and labeled with TRITC-phalloidin to visualize actin filaments.
Magnification 400x.