Supplemental Figure 4. Actin-Rich Cytoplasmic Clumps Are Generated in PalladinFPAA-Expressing Cells.

<p>HeLa cells were transfected with various GFP-tagged palladin DNA constructs (indicated on left), fixed and stained with DAPI (blue) to visualize DNA and Alexa594-phalloidin to visualize actin and simultaneously observe for general cytoskeletal alterations. </p> <p>(A) Palladin displays robust punctate staining at actin stress fibers in GFP-palladin transfected cells. In GFP-palladinFPAA cells, actin stress fibers are generally absent from within the central body of the cell and abnormal actin-rich cytoplasmic clumps (open arrowheads) and bar-like structures (solid arrowheads) are formed. Palladin retains some punctate staining at residual stress fibers within the cell. The actin cytoskeleton is disorganized and palladin is redistributed to the nucleus in GFP-palladinK15/18/51A transfected cells. Scale bar, 5 µm (also applies to B).</p> <p>(B) Endogenous VASP (red) is redistributed to actin-rich cytoplasmic clumps and bar-like structures formed in GFP-palladinFPAA transfected cells.</p>