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Fig. 6. XFlop depletion specifically affects the actin skeleton. (A) The
microtubule and intermediate filament networks are not significantly affected
by XFlop depletion. In a single experiment, control (upper row) and
XFlop-depleted (lower row; oligo 1s, 12 ng; oligo 5s, 10 ng) late blastulae
were divided into two groups (with five to six embryos in each). The first
group was assayed for F-actin by phalloidin staining. The second group was
fixed with FG fix with or without 0.5 µM Taxol for microtubule (MT)
staining or cytokeratin (CK) staining, respectively. Although significant
depletion of the F-actin network was seen, no significant defects in
microtubule or cytokeratin filament staining was seen. (B) The decrease of
F-actin observed in the experiment shown in A is significant
(P<0.001; five to six caps were scored in each case). (C) The
bases of control and XFlop-depleted embryos with or without injection of 100
pg XFlop mRNA into the animal cytoplasm at the two-cell stage.
XFlop mRNA rescues the wound healing and loss of rigidity caused by
the depletion of XFlop mRNA in the oocyte. (D) Cortical actin is also
rescued, as shown in animal caps from the same embryos stained with Alexa-488
phalloidin. (E) Quantitative analysis of the overall amount of the cortical
actin from the experiment shown in C and D (compare the control with the
XFlop-depleted animal caps, P<0.03; XFlop depletion with XFlop
mRNA rescue, P<0.001). The rescue experiments have been repeated
four times with the similar results. In each experiment, five to six caps from
each treatment were analyzed.
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