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Fig. 3. XFlop is sufficient for actin assembly in early Xenopus embryos.
(A) XFlop increases the rate of the wound healing in a dose-dependent manner.
Photographs were taken 45 minutes after the excision of animal caps from
embryos injected at the two-cell stage with 10-250 pg doses of XFlop
mRNA. (B) Injection of 100 pg of XFlop mRNA at the two-cell stage
(lower embryos) caused an increased thickness of the blastocoel roof when
compared with control embryos (upper) at stage 9. Embryos were fixed in FG fix
and split open for photography along the animal/vegetal axis. (C) The internal
(blastocoelic) surfaces of animal caps from stage 9 embryos injected at the
two-cell stage with 10-250 pg XFlop mRNA and cultured for ten minutes
before fixation and staining with Alexa-488 Phalloidin. XFlop increases
cortical actin assembly in a dose-dependent manner (arrow indicates a dividing
cell); shown are images of whole caps (upper row) and their pixel intensity
measurements (lower row). (D) Quantitative analysis of pixel intensities
measured from the experiment shown in C. Five to six caps for each treatment
were measured for pixel intensity quantitation. Student's t-test
shows that XFlop increases the overall amount of cortical actin significantly
in a dose-dependent manner (P<0.01 at all doses). (E) The
wound-healing rate of bases was significantly changed by either overexpressing
(P<0.01, yellow) or underexpressing (P<0.001, red)
XFlop. The width of the wound from each base was measured 1 hour after animal
cap removal. The ratio of the width of the wound to the diameter of the whole
embryo represents the wound healing rate (y axis, mean±s.d.).
Forty-four bases from control, 14 from Xflop-overexpressing and 33 from
Xflop-depleted embryos were scored.
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