Overexpression of nm23H1 reduces the adhesion-stimulated membrane
ruffles of Rat1 cells. (a–d) Rat1 cells were stained
for Tiam1 (a and c) and for F-actin
(b and d) on uncoated slides
(a and b) or replated on
fibronectin-coated slides for 15 min (c and
d). (e–j and l–o) Rat1
cells stably expressing GFP (e, g, i, l, and
n) or GFP-tagged nm23H1 (f, h, j,
m, and o) were either untransfected
(e–j) or were transfected with Tiam1 C1199 (myc tagged
at the C terminus) (l–o), then replated on
fibronectin-coated slides for 15 min and stained for F-actin (e,
f, l, and m), endogenous Tiam1
(g and h), or Tiam1 C1199 (anti-myc)
(n and o). The same cells are shown in
a–d, g, i, and h, j. (i
and j) Stable expression of GFP or GFP-nm23H1.
Expression of endogenous or transfected Tiam1 and nm23H1 in
Rat1 cells is shown by immunoblot (k and
p). Three independent clones expressing GFP-tagged
nm23H1 or GFP were examined, and similar results were obtained.