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Fig. 8. Electroporation of constitutively active RhoA or ROCK, or wild-type
ROCK, disrupts convergent extension in Lp/+ and +/+ embryos.
(A-F) Lp/+ and +/+ embryos after 18 hours culture following
electroporation in the node region with vector only (A), RhoA active (B,E),
ROCK active (C,F) or ROCK wild type (D). The vector control shows marked
midline extension of GFP-positive cells (arrows in A), by contrast to the
sparse, clumped appearance of cells electroporated with RhoA active (arrows in
B,E) or ROCK active (arrows in C,F). Electroporation with ROCK wild type
allows greater midline extension (arrows in D) although less than in the
vector control. (G,H) 293T cells transfected with RhoA active
(H) show reduced protrusive behaviour compared with vector-only controls (G).
(I) Immunoblot for phosphorylated cofilin in 293T cells transfected
with vector only (lane 1), RhoA active (lane 2), ROCK wild type (lane 3) or
ROCK active (lane 4). Beta tubulin (lower panel) serves as a loading control.
Note the severely reduced phosphocofilin abundance in cells expressing
constitutively active RhoA and mild downregulation of phosphocofilin in cells
expressing wild type and constitutively active ROCK. Scale bar: in F, 0.4 mm
for A-F.
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