Fig. 6. Effect of amotl2 mutants on cell migration and structures.
(A) Effect of overexpression of amotl2
PDZ
mutant on cell movements of zebrafish embryos. Embryos were injected at the
one-cell stage and observed at the indicated stages. Embryos were managed to
position with animal pole or anterior to the top. (B) Expression of
hgg1 (red), dlx3 (blue) and ntl (blue) at the
two-somite stage. Top panel, anterodorsal views; middle panel, dorsal views
with anterior to the top; bottom panel, lateral views with anterior to the top
and dorsal to the right. Injection of amotl2PDZ mRNA
led to varying degrees of defects in convergent extension (in the second and
third columns). Injection of amotl2EILI mRNA also
caused defective convergent extension (fourth column). The hgg1
expression domain is indicated by arrowheads. (C) Expression of
Amotl2PDZ-GFP or Amotl2-GFP inhibited or promoted cell migration in
vitro, respectively. Wound-healing assays were done both in HEK293T (shown on
the left) and in COS1 cells. The wound area was between two lines. The bar
graph at the right shows statistical data from three experiments with standard
deviations. The migration efficiency of GFP-positive cells was calculated as
percentage of GFP-positive cells in the wound-healing area/percentage of
GFP-positive cells in the non-wound area. (D) F-actin distribution was
abnormal in HeLa cells transfected with either Myc-Amotl2PDZ (derived
from fish) or HA-AMOTL2PDZ (derived from human). Cells were stained
with phalloidin 24 hours after transfection. Arrowheads indicate
Amotl2-expressing cells. Scale bars: 10 µm.