FIG. 2.

(A) Representative morphologies of N1E-115 cells. Gem stimulates cell flattening and neurite extension. (B) Effects of Gem and Gem mutants on neurite extension in N1E-115 murine neuroblastoma cells. Cells were cotransfected with pEGFP-N1 and Gem plasmids as indicated. The percentage of transfected cells with neurites is shown (from an average of five independent experiments) with standard deviations. Mutation of serine 261 or 289 to alanine inhibits the cytoskeletal effects of Gem. (C) Effects of Gem and Gem mutants on focal adhesion dissolution. HeLa cells were transfected with empty vector and pEGFP-N1 or various Gem cDNAs. Gem-expressing cells were stained with anti-Gem polyclonal antibody followed by an anti-rabbit fluorescein isothiocyanate-conjugated antibody and antivinculin monoclonal antibody followed by an anti-mouse rhodamine-conjugated secondary antibody. Cells expressing empty vector and GFP were stained for vinculin only. The percent transfected cells containing focal adhesions throughout the cell surface was quantified in three independent experiments. Four hundred cells were counted for each. (D) Confocal images showing representative focal adhesion patterns.